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e0554s camp glosensor assay promega  (New England Biolabs)


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    New England Biolabs e0554s camp glosensor assay promega
    E0554s Camp Glosensor Assay Promega, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 5863 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/glosensor+kit/Q5+Site-Directed+Mutagenesis+Kit/pm33571432-397-127-125
    Average 99 stars, based on 5863 article reviews
    e0554s camp glosensor assay promega - by Bioz Stars, 2026-11
    99/100 stars

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    Related Articles

    Mutagenesis:

    Article Title: A Blast Fungus Effector Targets a Tubby-like F-box Protein to Suppress Plant Immunity
    Article Snippet: .. Site directed mutagenesis was performed using the Q5 Site-Directed Mutagenesis Kit from New England Biolabs (Ipswich, MA, USA) according to the manufacturers protocol. ..

    Article Title: BATF condensation drives gastric cancer metastasis via chromatin association and transcriptional regulation.
    Article Snippet: .. Mutagenesis was performed using the Q5 Site-Directed Mutagenesis Kit (NEB, E0554S) according to the manufacturer’s instructions. ..

    Article Title: Chemogenetic ubiquitin E1 inhibitor TAK-243 CRISPR screen uncovers HAPSTR1/HUWE1 axis in chromatin proteostasis and cell cycle control
    Article Snippet: The cDNA of HAPSTR1 was obtained from the MISSION TRC Human ORF Collection (Sigma-Aldrich, clone ID 03065) in the donor vector pDONR223. .. HAPSTR1-ΔA96 was generated via site-directed mutagenesis using the Q5 site-directed mutagenesis kit according to the manufacturer’s protocol (NEB, #E0554S). ..

    Article Title: Molecular triage and isoform protection safeguard ubiquitin chain editing on DNA.
    Article Snippet: All plasmids were generated by using either PCR amplification with Q5 DNA polymerase (New England Biolabs, NEB), HiFi assembly (NEB), or restriction enzyme digestion followed by ligation, or some combination of these, using plasmid DNA, gBlocks (IDT), or synthesized genes (Thermo Fisher Scientific) as templates. .. Site-directed mutagenesis was carried out using the Q5 DNA polymerase and KLD enzyme mix (NEB). .. All plasmid DNA was purified from the appropriate E. coli strains using either Miniprep (Qiagen) or PureLink HiPure Plasmid Filter Maxiprep Kit (Thermo Fisher Scientific) kits, respectively.

    Article Title: Tryptoline stereoprobe elaboration identifies inhibitors of the GRPEL1-HSPA9 complex.
    Article Snippet: All full-length plasmids were obtained from either OriGene in a pCMV6 vector with a C-terminal Myc-DDK (Flag) epitope tag or from GenScript in pcDNA3.1-C-(k) DYK (Flag), as shown in Supplementary Table 2. .. Mutagenesis was performed using a Q5 site-directed mutagenesis kit (New England BioLabs, E0554S) and the primers shown in Supplementary Table 3. ..

    Article Title: Discovery of direct-acting small-molecule RIG-I agonists through creation of a selective CARD activation platform
    Article Snippet: .. The RIG-I constructs used in this study were 3G-DC-RIG-I and3G-DC-527TAG-RIG-I, all of which were mutated from wild-type human RIG-I in a pET SUMO expression vector using the Q5 Site-Directed Mutagenesis Kit (NEB). ..

    Site-directed Mutagenesis:

    Article Title: BATF condensation drives gastric cancer metastasis via chromatin association and transcriptional regulation.
    Article Snippet: .. Mutagenesis was performed using the Q5 Site-Directed Mutagenesis Kit (NEB, E0554S) according to the manufacturer’s instructions. ..

    Article Title: Tryptoline stereoprobe elaboration identifies inhibitors of the GRPEL1-HSPA9 complex.
    Article Snippet: All full-length plasmids were obtained from either OriGene in a pCMV6 vector with a C-terminal Myc-DDK (Flag) epitope tag or from GenScript in pcDNA3.1-C-(k) DYK (Flag), as shown in Supplementary Table 2. .. Mutagenesis was performed using a Q5 site-directed mutagenesis kit (New England BioLabs, E0554S) and the primers shown in Supplementary Table 3. ..

    Article Title: Osteopontin induces cyclo-oxygenase-2 expression in osteoclasts.
    Article Snippet: Luminescence was measured using a TD-20/20 luminometer (Turner Designs, Inc. San Jose, CA, USA) as relative light units (RLUs). .. Site-directed mutagenesis was performed using the Q5 Site-Directed Mutagenesis Kit (New England Biolabs, Ipswich, MA) following the supplied instructions. ..

    Article Title: Molecular mechanism of residue-specific nucleosome acetylation.
    Article Snippet: Reshuffled sequences were designed using the SeqMixer application and ordered as gBlocks from Integrated DNA Technologies. .. Point mutations or deletions were introduced using the Q5 site-directed mutagenesis kit (New England Biolabs). ..

    Generated:

    Article Title: Chemogenetic ubiquitin E1 inhibitor TAK-243 CRISPR screen uncovers HAPSTR1/HUWE1 axis in chromatin proteostasis and cell cycle control
    Article Snippet: The cDNA of HAPSTR1 was obtained from the MISSION TRC Human ORF Collection (Sigma-Aldrich, clone ID 03065) in the donor vector pDONR223. .. HAPSTR1-ΔA96 was generated via site-directed mutagenesis using the Q5 site-directed mutagenesis kit according to the manufacturer’s protocol (NEB, #E0554S). ..

    Construct:

    Article Title: Discovery of direct-acting small-molecule RIG-I agonists through creation of a selective CARD activation platform
    Article Snippet: .. The RIG-I constructs used in this study were 3G-DC-RIG-I and3G-DC-527TAG-RIG-I, all of which were mutated from wild-type human RIG-I in a pET SUMO expression vector using the Q5 Site-Directed Mutagenesis Kit (NEB). ..

    Expressing:

    Article Title: Discovery of direct-acting small-molecule RIG-I agonists through creation of a selective CARD activation platform
    Article Snippet: .. The RIG-I constructs used in this study were 3G-DC-RIG-I and3G-DC-527TAG-RIG-I, all of which were mutated from wild-type human RIG-I in a pET SUMO expression vector using the Q5 Site-Directed Mutagenesis Kit (NEB). ..

    Plasmid Preparation:

    Article Title: Discovery of direct-acting small-molecule RIG-I agonists through creation of a selective CARD activation platform
    Article Snippet: .. The RIG-I constructs used in this study were 3G-DC-RIG-I and3G-DC-527TAG-RIG-I, all of which were mutated from wild-type human RIG-I in a pET SUMO expression vector using the Q5 Site-Directed Mutagenesis Kit (NEB). ..



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    FUT8 knockdown attenuates the FSH/FSHR signaling pathway. A, CCK8 cell proliferation assay. Cells were treated with 100 ng/mL rhFSH, and cell proliferation was measured by absorbance at a wavelength of 450 nm. Significant differences are shown as the means ± SD, n = 4. B, Immunofluorescence assay. KGN, KGN-KD and KGN-KD-Re cells were treated with FSH (400 ng/mL) and stained with anti-FSHR (red) (1:500) and DAPI (blue). C, Western blot. Cell lysates of KGN, KGN-KD and KGN-KD-Re cells were resolved by 7.5 % SDS-PAGE, transferred to a PVDF membrane and probed with anti-FSHR (1:2000). GAPDH was used as the reference gene for the cytoplasm. D, Western blot. Cell lysates of Fut8 +/+ and Fut8 −/− ovaries were resolved by 7.5 % SDS-PAGE, transferred to a PVDF membrane and probed with anti-FSHR (1:2000). GAPDH was used as a loading control. E, Quantitative real-time PCR. RNA was isolated from the granulosa cells of MOR and POR patients. The mRNA expression levels of FSHR were compared. The relative expression of mRNAs was normalized to GAPDH. Data are presented as the means ± SEM, n = 3. F, FSH-induced cAMP production was measured using a <t>GloSensor</t> cAMP Assay kit. Cells were seeded in a 6-well plate for 24 h, and then 100 μM of IBMX was added, to inhibit phosphodiesterases, for 20 min. The cAMP levels were measured in KGN, KGN-KD, and KGN-KD-Re cells following stimulation with 100 IU/L of rhFSH or 100 μM of forskolin for 5 min. Data are presented as the means ± SEM, n = 3. G, PKA-CREB signaling pathway and Akt-FoxO3a signaling pathway. Serum-starved KGN, KGN-KD, and KGN-KD-Re cells were treated with FSH for 30 min, and cell lysates were resolved by 10 % SDS-PAGE, followed by transfer to a PVDF membrane. Membranes were probed with anti-phospho-PKA (1:2000) and anti-PKA (1:2000); anti-phospho-CREB (1:2000) and anti-CREB (1:2000); and anti-KITL (1:2000); anti-phospho-Akt (1:2000) and anti-Akt (1:2000); anti-phospho-FoxO3a (1:2000) and anti-FoxO3a (1:2000), and GAPDH was used as a loading control. Quantitative data are represented as the means ± SD, n = 3. H, Western blot. Lysates of Fut8 +/+ and Fut8 −/− ovaries were resolved by 10 % SDS-PAGE, and probed with anti-KITL (1:2000), with GAPDH used as the loading control. Quantitative data are represented as the means ± SD, n = 3. I, Western blot. Lysates of Fut8 +/+ and Fut8 −/− ovaries were resolved by 10 % SDS-PAGE and probed with anti-pFoxO3a (1:2000) and anti-FoxO3a (1:2000). Quantitative data are represented as the means ± SD, n = 3. J, TUNEL assay in the Fut8 +/+ and Fut8 −/− ovarian follicles. Representative micrographs are shown. Quantitative data are represented as the means ± SD, n = 3. K, Western blot analysis. Cell lysates of Fut8 +/+ and Fut8 −/− ovaries were resolved by 7.5 % SDS-PAGE, transferred to a PVDF membrane, and probed with anti-BCL2 and anti-BAX (1:2000). Quantitative data are represented as the means ± SD, n = 3. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
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    FUT8 knockdown attenuates the FSH/FSHR signaling pathway. A, CCK8 cell proliferation assay. Cells were treated with 100 ng/mL rhFSH, and cell proliferation was measured by absorbance at a wavelength of 450 nm. Significant differences are shown as the means ± SD, n = 4. B, Immunofluorescence assay. KGN, KGN-KD and KGN-KD-Re cells were treated with FSH (400 ng/mL) and stained with anti-FSHR (red) (1:500) and DAPI (blue). C, Western blot. Cell lysates of KGN, KGN-KD and KGN-KD-Re cells were resolved by 7.5 % SDS-PAGE, transferred to a PVDF membrane and probed with anti-FSHR (1:2000). GAPDH was used as the reference gene for the cytoplasm. D, Western blot. Cell lysates of Fut8 +/+ and Fut8 −/− ovaries were resolved by 7.5 % SDS-PAGE, transferred to a PVDF membrane and probed with anti-FSHR (1:2000). GAPDH was used as a loading control. E, Quantitative real-time PCR. RNA was isolated from the granulosa cells of MOR and POR patients. The mRNA expression levels of FSHR were compared. The relative expression of mRNAs was normalized to GAPDH. Data are presented as the means ± SEM, n = 3. F, FSH-induced cAMP production was measured using a GloSensor cAMP Assay kit. Cells were seeded in a 6-well plate for 24 h, and then 100 μM of IBMX was added, to inhibit phosphodiesterases, for 20 min. The cAMP levels were measured in KGN, KGN-KD, and KGN-KD-Re cells following stimulation with 100 IU/L of rhFSH or 100 μM of forskolin for 5 min. Data are presented as the means ± SEM, n = 3. G, PKA-CREB signaling pathway and Akt-FoxO3a signaling pathway. Serum-starved KGN, KGN-KD, and KGN-KD-Re cells were treated with FSH for 30 min, and cell lysates were resolved by 10 % SDS-PAGE, followed by transfer to a PVDF membrane. Membranes were probed with anti-phospho-PKA (1:2000) and anti-PKA (1:2000); anti-phospho-CREB (1:2000) and anti-CREB (1:2000); and anti-KITL (1:2000); anti-phospho-Akt (1:2000) and anti-Akt (1:2000); anti-phospho-FoxO3a (1:2000) and anti-FoxO3a (1:2000), and GAPDH was used as a loading control. Quantitative data are represented as the means ± SD, n = 3. H, Western blot. Lysates of Fut8 +/+ and Fut8 −/− ovaries were resolved by 10 % SDS-PAGE, and probed with anti-KITL (1:2000), with GAPDH used as the loading control. Quantitative data are represented as the means ± SD, n = 3. I, Western blot. Lysates of Fut8 +/+ and Fut8 −/− ovaries were resolved by 10 % SDS-PAGE and probed with anti-pFoxO3a (1:2000) and anti-FoxO3a (1:2000). Quantitative data are represented as the means ± SD, n = 3. J, TUNEL assay in the Fut8 +/+ and Fut8 −/− ovarian follicles. Representative micrographs are shown. Quantitative data are represented as the means ± SD, n = 3. K, Western blot analysis. Cell lysates of Fut8 +/+ and Fut8 −/− ovaries were resolved by 7.5 % SDS-PAGE, transferred to a PVDF membrane, and probed with anti-BCL2 and anti-BAX (1:2000). Quantitative data are represented as the means ± SD, n = 3. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Journal: Journal of Advanced Research

    Article Title: Core fucosylation regulates the ovarian response via FSH receptor during follicular development

    doi: 10.1016/j.jare.2024.01.025

    Figure Lengend Snippet: FUT8 knockdown attenuates the FSH/FSHR signaling pathway. A, CCK8 cell proliferation assay. Cells were treated with 100 ng/mL rhFSH, and cell proliferation was measured by absorbance at a wavelength of 450 nm. Significant differences are shown as the means ± SD, n = 4. B, Immunofluorescence assay. KGN, KGN-KD and KGN-KD-Re cells were treated with FSH (400 ng/mL) and stained with anti-FSHR (red) (1:500) and DAPI (blue). C, Western blot. Cell lysates of KGN, KGN-KD and KGN-KD-Re cells were resolved by 7.5 % SDS-PAGE, transferred to a PVDF membrane and probed with anti-FSHR (1:2000). GAPDH was used as the reference gene for the cytoplasm. D, Western blot. Cell lysates of Fut8 +/+ and Fut8 −/− ovaries were resolved by 7.5 % SDS-PAGE, transferred to a PVDF membrane and probed with anti-FSHR (1:2000). GAPDH was used as a loading control. E, Quantitative real-time PCR. RNA was isolated from the granulosa cells of MOR and POR patients. The mRNA expression levels of FSHR were compared. The relative expression of mRNAs was normalized to GAPDH. Data are presented as the means ± SEM, n = 3. F, FSH-induced cAMP production was measured using a GloSensor cAMP Assay kit. Cells were seeded in a 6-well plate for 24 h, and then 100 μM of IBMX was added, to inhibit phosphodiesterases, for 20 min. The cAMP levels were measured in KGN, KGN-KD, and KGN-KD-Re cells following stimulation with 100 IU/L of rhFSH or 100 μM of forskolin for 5 min. Data are presented as the means ± SEM, n = 3. G, PKA-CREB signaling pathway and Akt-FoxO3a signaling pathway. Serum-starved KGN, KGN-KD, and KGN-KD-Re cells were treated with FSH for 30 min, and cell lysates were resolved by 10 % SDS-PAGE, followed by transfer to a PVDF membrane. Membranes were probed with anti-phospho-PKA (1:2000) and anti-PKA (1:2000); anti-phospho-CREB (1:2000) and anti-CREB (1:2000); and anti-KITL (1:2000); anti-phospho-Akt (1:2000) and anti-Akt (1:2000); anti-phospho-FoxO3a (1:2000) and anti-FoxO3a (1:2000), and GAPDH was used as a loading control. Quantitative data are represented as the means ± SD, n = 3. H, Western blot. Lysates of Fut8 +/+ and Fut8 −/− ovaries were resolved by 10 % SDS-PAGE, and probed with anti-KITL (1:2000), with GAPDH used as the loading control. Quantitative data are represented as the means ± SD, n = 3. I, Western blot. Lysates of Fut8 +/+ and Fut8 −/− ovaries were resolved by 10 % SDS-PAGE and probed with anti-pFoxO3a (1:2000) and anti-FoxO3a (1:2000). Quantitative data are represented as the means ± SD, n = 3. J, TUNEL assay in the Fut8 +/+ and Fut8 −/− ovarian follicles. Representative micrographs are shown. Quantitative data are represented as the means ± SD, n = 3. K, Western blot analysis. Cell lysates of Fut8 +/+ and Fut8 −/− ovaries were resolved by 7.5 % SDS-PAGE, transferred to a PVDF membrane, and probed with anti-BCL2 and anti-BAX (1:2000). Quantitative data are represented as the means ± SD, n = 3. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

    Article Snippet: FSH-induced cAMP production was measured using a GloSensor cAMP Assay kit (Promega).

    Techniques: Knockdown, Proliferation Assay, Immunofluorescence, Staining, Western Blot, SDS Page, Membrane, Control, Real-time Polymerase Chain Reaction, Isolation, Expressing, cAMP Assay, TUNEL Assay