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alphalisa readings  (BMG Labtech)


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    Structured Review

    BMG Labtech alphalisa readings
    G protein-coupled receptor 35 (GPR35) activation reduces c-Src and PI3K-Akt signaling pathways in mature osteoclasts. Quantification of phosphorylated (A) c-Src (pSrc), (B) Akt1/2/3 (pAkt), and (C) p38 (p-p38) measured by <t>AlphaLISA</t> in osteoclasts exposed to vehicle or GPR35 agonists with or without GPR35 antagonist ML145. (D-E) Quantification of (D) c-Src and (E) Akt1/2/3 phosphorylation in mature osteoclasts transfected with scrambled or GPR35 siRNA exposed to vehicle or GPR35 agonist, TC-G 1001. Data in all panels was normalized to GAPDH as a housekeeper control. Each point represents an independent donor in all panels. The gray line denotes mean in panels a, B, D and E and median in panel C. Statistical analyses were performed by one-way ANOVA with Holm-Šídák’s multiple comparisons test in panels A, B, D, and E and Kruskal–Wallis with Dunn’s multiple comparisons test in C. **** p < .0001, *** p < .001, ** p < .01, * p < .05.
    Alphalisa Readings, supplied by BMG Labtech, used in various techniques. Bioz Stars score: 99/100, based on 4228 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/alphalisa+readings/PHERAstar+FSX/pmc12445851-62-0-8
    Average 99 stars, based on 4228 article reviews
    alphalisa readings - by Bioz Stars, 2026-10
    99/100 stars

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    1) Product Images from "G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells"

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells

    Journal: JBMR Plus

    doi: 10.1093/jbmrpl/ziaf131

    G protein-coupled receptor 35 (GPR35) activation reduces c-Src and PI3K-Akt signaling pathways in mature osteoclasts. Quantification of phosphorylated (A) c-Src (pSrc), (B) Akt1/2/3 (pAkt), and (C) p38 (p-p38) measured by AlphaLISA in osteoclasts exposed to vehicle or GPR35 agonists with or without GPR35 antagonist ML145. (D-E) Quantification of (D) c-Src and (E) Akt1/2/3 phosphorylation in mature osteoclasts transfected with scrambled or GPR35 siRNA exposed to vehicle or GPR35 agonist, TC-G 1001. Data in all panels was normalized to GAPDH as a housekeeper control. Each point represents an independent donor in all panels. The gray line denotes mean in panels a, B, D and E and median in panel C. Statistical analyses were performed by one-way ANOVA with Holm-Šídák’s multiple comparisons test in panels A, B, D, and E and Kruskal–Wallis with Dunn’s multiple comparisons test in C. **** p < .0001, *** p < .001, ** p < .01, * p < .05.
    Figure Legend Snippet: G protein-coupled receptor 35 (GPR35) activation reduces c-Src and PI3K-Akt signaling pathways in mature osteoclasts. Quantification of phosphorylated (A) c-Src (pSrc), (B) Akt1/2/3 (pAkt), and (C) p38 (p-p38) measured by AlphaLISA in osteoclasts exposed to vehicle or GPR35 agonists with or without GPR35 antagonist ML145. (D-E) Quantification of (D) c-Src and (E) Akt1/2/3 phosphorylation in mature osteoclasts transfected with scrambled or GPR35 siRNA exposed to vehicle or GPR35 agonist, TC-G 1001. Data in all panels was normalized to GAPDH as a housekeeper control. Each point represents an independent donor in all panels. The gray line denotes mean in panels a, B, D and E and median in panel C. Statistical analyses were performed by one-way ANOVA with Holm-Šídák’s multiple comparisons test in panels A, B, D, and E and Kruskal–Wallis with Dunn’s multiple comparisons test in C. **** p < .0001, *** p < .001, ** p < .01, * p < .05.

    Techniques Used: Activation Assay, Protein-Protein interactions, Phospho-proteomics, Transfection, Control

    G protein-coupled receptor 35 (GPR35) mediated effects on bone resorption and signaling are maintained in osteoclast-osteoblast co-cultures. (A) Quantification of toluidine blue-stained resorption pits and trenches formed on bone slices by osteoclast-osteoblast co-cultures exposed to vehicle, TC-G 1001, or TC-G 1001 and the ML145 antagonist (B) quantification of tartrate-resistant acid phosphatase enzyme activity in conditioned media from osteoclast-osteoblast co-cultures exposed to GPR35 agonists or agonists with antagonist. (C-F) quantification of (C) phosphorylated c-Src, (D) phosphorylated Akt1/2/3, (E) phosphorylated NFκB, (F) phosphorylated cAMP response element-binding protein in osteoclast-osteoblast co-cultures exposed to GPR35 agonists or agonists with antagonist, measured by AlphaLISA. Each point represents an independent donor. The gray line denotes mean. Statistical analyses were performed by one-way ANOVA with Holm-Šídák’s multiple comparisons test for A-B and with Dunnett’s multiple comparisons test for C-F. **** p < .0001, *** p < .001, ** p < .01, * p < .05.
    Figure Legend Snippet: G protein-coupled receptor 35 (GPR35) mediated effects on bone resorption and signaling are maintained in osteoclast-osteoblast co-cultures. (A) Quantification of toluidine blue-stained resorption pits and trenches formed on bone slices by osteoclast-osteoblast co-cultures exposed to vehicle, TC-G 1001, or TC-G 1001 and the ML145 antagonist (B) quantification of tartrate-resistant acid phosphatase enzyme activity in conditioned media from osteoclast-osteoblast co-cultures exposed to GPR35 agonists or agonists with antagonist. (C-F) quantification of (C) phosphorylated c-Src, (D) phosphorylated Akt1/2/3, (E) phosphorylated NFκB, (F) phosphorylated cAMP response element-binding protein in osteoclast-osteoblast co-cultures exposed to GPR35 agonists or agonists with antagonist, measured by AlphaLISA. Each point represents an independent donor. The gray line denotes mean. Statistical analyses were performed by one-way ANOVA with Holm-Šídák’s multiple comparisons test for A-B and with Dunnett’s multiple comparisons test for C-F. **** p < .0001, *** p < .001, ** p < .01, * p < .05.

    Techniques Used: Staining, Activity Assay, Binding Assay

    Related Articles

    Activation Assay:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Protein-Protein interactions:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Phospho-proteomics:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Transfection:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Control:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Staining:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Activity Assay:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Binding Assay:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Variant Assay:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Expressing:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Concentration Assay:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Luciferase:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Generated:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Fluorescence:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Imaging:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    MANN-WHITNEY:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Western Blot:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    Inhibition:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.

    TUNEL Assay:

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), c

    Article Title: Characterisation of a GNAS variant linked to cortisol-producing adrenocortical adenoma
    Article Snippet: Revvity, UK) and pCREB and GAPDH (Catalog# ALSU-TGAPD-A500, Revvity, Oxford) assays performed according to the manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, UK) plate reader in duplicates and values for phosphorylated proteins normalized to GAPDH values. .. Ratios of pCREB:GAPDH were expressed as a percentage of the wild-type Gα s E max and dose–response curves fitted with a three-parameter log agonist cu

    Article Title: G protein-coupled receptor 35 (GPR35) stimulation reduces osteoclast activity in primary human bone cells
    Article Snippet: were transferred to white 384-well plates (OptiPlates, PerkinElmer) in duplicates and assays performed according to manufacturer’s instructions. .. AlphaLISA readings were made on a Pherastar FS (BMG Labtech, Aylesbury, UK) plate reader and values for phosphorylated proteins normalized to GAPDH values. .. The following AlphaLISA assay kits (PerkinElmer) were performed according to manufacturer’s instructions: phosphorylated forms of Akt1/2/3 (Ser473), C

    Article Title: GIP receptor reduces osteoclast activity and improves osteoblast survival by activating multiple signaling pathways
    Article Snippet: Akt1/2/3 (Ser473), p65 NFkB (Ser536) and p38 (Thr180/Thr182) were purchased from PerkinElmer and performed according to manufacturer’s instructions. .. Assays were performed in white 384-well plates (Optiplates, PerkinElmer) and AlphaLISA readings made on a Pherastar FS (BMG Labtech) plate reader. .. Values for phosphorylated proteins were normalized to GAPDH values.



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    BMG Labtech alphalisa readings
    G protein-coupled receptor 35 (GPR35) activation reduces c-Src and PI3K-Akt signaling pathways in mature osteoclasts. Quantification of phosphorylated (A) c-Src (pSrc), (B) Akt1/2/3 (pAkt), and (C) p38 (p-p38) measured by <t>AlphaLISA</t> in osteoclasts exposed to vehicle or GPR35 agonists with or without GPR35 antagonist ML145. (D-E) Quantification of (D) c-Src and (E) Akt1/2/3 phosphorylation in mature osteoclasts transfected with scrambled or GPR35 siRNA exposed to vehicle or GPR35 agonist, TC-G 1001. Data in all panels was normalized to GAPDH as a housekeeper control. Each point represents an independent donor in all panels. The gray line denotes mean in panels a, B, D and E and median in panel C. Statistical analyses were performed by one-way ANOVA with Holm-Šídák’s multiple comparisons test in panels A, B, D, and E and Kruskal–Wallis with Dunn’s multiple comparisons test in C. **** p < .0001, *** p < .001, ** p < .01, * p < .05.
    Alphalisa Readings, supplied by BMG Labtech, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    G protein-coupled receptor 35 (GPR35) activation reduces c-Src and PI3K-Akt signaling pathways in mature osteoclasts. Quantification of phosphorylated (A) c-Src (pSrc), (B) Akt1/2/3 (pAkt), and (C) p38 (p-p38) measured by AlphaLISA in osteoclasts exposed to vehicle or GPR35 agonists with or without GPR35 antagonist ML145. (D-E) Quantification of (D) c-Src and (E) Akt1/2/3 phosphorylation in mature osteoclasts transfected with scrambled or GPR35 siRNA exposed to vehicle or GPR35 agonist, TC-G 1001. Data in all panels was normalized to GAPDH as a housekeeper control. Each point represents an independent donor in all panels. The gray line denotes mean in panels a, B, D and E and median in panel C. Statistical analyses were performed by one-way ANOVA with Holm-Šídák’s multiple comparisons test in panels A, B, D, and E and Kruskal–Wallis with Dunn’s multiple comparisons test in C. **** p < .0001, *** p < .001, ** p < .01, * p < .05.

    Journal: JBMR Plus

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells

    doi: 10.1093/jbmrpl/ziaf131

    Figure Lengend Snippet: G protein-coupled receptor 35 (GPR35) activation reduces c-Src and PI3K-Akt signaling pathways in mature osteoclasts. Quantification of phosphorylated (A) c-Src (pSrc), (B) Akt1/2/3 (pAkt), and (C) p38 (p-p38) measured by AlphaLISA in osteoclasts exposed to vehicle or GPR35 agonists with or without GPR35 antagonist ML145. (D-E) Quantification of (D) c-Src and (E) Akt1/2/3 phosphorylation in mature osteoclasts transfected with scrambled or GPR35 siRNA exposed to vehicle or GPR35 agonist, TC-G 1001. Data in all panels was normalized to GAPDH as a housekeeper control. Each point represents an independent donor in all panels. The gray line denotes mean in panels a, B, D and E and median in panel C. Statistical analyses were performed by one-way ANOVA with Holm-Šídák’s multiple comparisons test in panels A, B, D, and E and Kruskal–Wallis with Dunn’s multiple comparisons test in C. **** p < .0001, *** p < .001, ** p < .01, * p < .05.

    Article Snippet: AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values.

    Techniques: Activation Assay, Protein-Protein interactions, Phospho-proteomics, Transfection, Control

    G protein-coupled receptor 35 (GPR35) mediated effects on bone resorption and signaling are maintained in osteoclast-osteoblast co-cultures. (A) Quantification of toluidine blue-stained resorption pits and trenches formed on bone slices by osteoclast-osteoblast co-cultures exposed to vehicle, TC-G 1001, or TC-G 1001 and the ML145 antagonist (B) quantification of tartrate-resistant acid phosphatase enzyme activity in conditioned media from osteoclast-osteoblast co-cultures exposed to GPR35 agonists or agonists with antagonist. (C-F) quantification of (C) phosphorylated c-Src, (D) phosphorylated Akt1/2/3, (E) phosphorylated NFκB, (F) phosphorylated cAMP response element-binding protein in osteoclast-osteoblast co-cultures exposed to GPR35 agonists or agonists with antagonist, measured by AlphaLISA. Each point represents an independent donor. The gray line denotes mean. Statistical analyses were performed by one-way ANOVA with Holm-Šídák’s multiple comparisons test for A-B and with Dunnett’s multiple comparisons test for C-F. **** p < .0001, *** p < .001, ** p < .01, * p < .05.

    Journal: JBMR Plus

    Article Title: G protein-coupled receptor 35 stimulation reduces osteoclast activity in primary human bone cells

    doi: 10.1093/jbmrpl/ziaf131

    Figure Lengend Snippet: G protein-coupled receptor 35 (GPR35) mediated effects on bone resorption and signaling are maintained in osteoclast-osteoblast co-cultures. (A) Quantification of toluidine blue-stained resorption pits and trenches formed on bone slices by osteoclast-osteoblast co-cultures exposed to vehicle, TC-G 1001, or TC-G 1001 and the ML145 antagonist (B) quantification of tartrate-resistant acid phosphatase enzyme activity in conditioned media from osteoclast-osteoblast co-cultures exposed to GPR35 agonists or agonists with antagonist. (C-F) quantification of (C) phosphorylated c-Src, (D) phosphorylated Akt1/2/3, (E) phosphorylated NFκB, (F) phosphorylated cAMP response element-binding protein in osteoclast-osteoblast co-cultures exposed to GPR35 agonists or agonists with antagonist, measured by AlphaLISA. Each point represents an independent donor. The gray line denotes mean. Statistical analyses were performed by one-way ANOVA with Holm-Šídák’s multiple comparisons test for A-B and with Dunnett’s multiple comparisons test for C-F. **** p < .0001, *** p < .001, ** p < .01, * p < .05.

    Article Snippet: AlphaLISA readings were made on a PHERAstar FS (BMG Labtech, Aylesbury, United Kingdom) plate reader, and values for phosphorylated proteins normalized to Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) values.

    Techniques: Staining, Activity Assay, Binding Assay