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pp3 #529574  (Millipore)


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    Structured Review

    Millipore pp3 #529574
    Pp3 #529574, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pp3/pp3++529574/pm40537792-343-3-8
    Average 90 stars, based on 1 article reviews
    pp3 #529574 - by Bioz Stars, 2026-11
    90/100 stars

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    Related Articles

    Protease Inhibitor:

    Article Title: Lemongrass essential oil and citral inhibit Src/Stat3 activity and suppress the proliferation/survival of small-cell lung cancer cells, alone or in combination with chemotherapeutic agents.
    Article Snippet: LG-EO and citral were dissolved in dimethyl sulfoxide (DMSO; Wako Pure Chemical Industries, Ltd., Osaka, Japan) at a concentration of 1% (v/v) before use. .. PP2 and PP3 dissolved in DMSO, the protease inhibitor cocktail set 1 and G418 were purchased from Calbiochem (San Diego, CA, USA). .. WST-1 was obtained from Roche Diagnostics (Mannheim, Germany).

    Article Title: Cbl-associated protein is tyrosine phosphorylated by c-Abl and c-Src kinases
    Article Snippet: .. Src inhibition was accomplished by incubating the cells with 5 μM PP1, PP2 or PP3 (Calbiochem) for 25 min. For detection of phosphorylated proteins, cells were lyzed in lysis buffer (100 mM Tris pH 8.0, 150 mM NaCl, 1 mM MgCl 2 , 1% Triton X-100, 60 mM n-Octylglucoside), supplemented with Protease Inhibitor Cocktail (Sigma) and 1 mM vanadate. .. Lysates were incubated at 4°C for 16 h with Protein A Dynabeads (Invitrogen) precoupled with the antibody.

    Incubation:

    Article Title: Dietary potassium stimulates Ppp1Ca-Ppp1r1a dephosphorylation of kidney NaCl cotransporter and reduces blood pressure
    Article Snippet: Excess FLAG peptides were removed from the IP eluate using Vivacon 500 spin columns (30,000 molecular weight cutoff [MWCO], Viva Products). .. Eluate was incubated with either (a) 10 units of Protein Phosphatase 1 Catalytic Subunit, α-isoform (MilliporeSigma, catalog P7937; 6123.08 units/mg) and 1 mM MnCl 2 in 1× PP1 reaction buffer (10 mM NaCl, 5 mM imidazole [pH 7.4], 0.2 mM DTT, 2.5‰Tween 20); (b) 25 ng PP2Aα (SignalChem, catalog P16-20BH) in 1× PP2 reaction buffer (25 mM HEPES [pH 7.2], 50 mM NaCl, 2.5 mM EDTA, 50 mM imidazole, 0.2% 2-mercaptolethanol, 65 ng/μL BSA); or (c) 10 units of PP3 (MilliporeSigma, catalog C1907), 1 mM MnCl 2 and 10 μg/mL calmodulin (MilliporeSigma, catalog P0270) in 1× PP3 reaction buffer (50 mM Tris-HCl [pH 7.0], 50 μM CaCl 2 , 50 μg/mL BSA) at 30°C for 30 minutes. ..

    other:

    Article Title: Bacillus anthracis Spore Entry into Epithelial Cells Is an Actin-Dependent Process Requiring c-Src and PI3K
    Article Snippet: Murine embryonic fibroblasts (MEFs) from src-/-yes-/-fyn-/- triple knockout mice were obtained from ATCC and cultured according to instructions from ATCC.

    Article Title: <scp>TRPV4</scp> mediates <scp>IL</scp>‐1‐induced Ca<sup>2+</sup> signaling, <scp>ERK</scp> activation and <scp>MMP</scp> expression
    Article Snippet: Funding information Canadian Government | Canadian Institutes of Health Research (CIHR), Grant/Award Number: MOP512788 Abstract Ca2+ permeation through TRPV4 in fibroblasts is associated with pathological matrix degradation.. In human gingival fibroblasts, IL1β binding to its signaling receptor (IL1R1) induces activation of extracellular regulated kinase (ERK) and MMP1 expression, processes that require Ca2+ flux across the plasma membrane.. It is not known how IL1R1, which does not conduct Ca2+, generates Ca2+ signals in response to IL1.

    Inhibition:

    Article Title: Cbl-associated protein is tyrosine phosphorylated by c-Abl and c-Src kinases
    Article Snippet: .. Src inhibition was accomplished by incubating the cells with 5 μM PP1, PP2 or PP3 (Calbiochem) for 25 min. For detection of phosphorylated proteins, cells were lyzed in lysis buffer (100 mM Tris pH 8.0, 150 mM NaCl, 1 mM MgCl 2 , 1% Triton X-100, 60 mM n-Octylglucoside), supplemented with Protease Inhibitor Cocktail (Sigma) and 1 mM vanadate. .. Lysates were incubated at 4°C for 16 h with Protein A Dynabeads (Invitrogen) precoupled with the antibody.

    Lysis:

    Article Title: Cbl-associated protein is tyrosine phosphorylated by c-Abl and c-Src kinases
    Article Snippet: .. Src inhibition was accomplished by incubating the cells with 5 μM PP1, PP2 or PP3 (Calbiochem) for 25 min. For detection of phosphorylated proteins, cells were lyzed in lysis buffer (100 mM Tris pH 8.0, 150 mM NaCl, 1 mM MgCl 2 , 1% Triton X-100, 60 mM n-Octylglucoside), supplemented with Protease Inhibitor Cocktail (Sigma) and 1 mM vanadate. .. Lysates were incubated at 4°C for 16 h with Protein A Dynabeads (Invitrogen) precoupled with the antibody.



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    Concentration-response relationships of ( A ) DAMGO as a recruiter of β-arrestin2 to human μ receptors in PathHunter CHO cells and ( B ) DAMGO as an inhibitor of forskolin (30 μM) stimulated cAMP accumulation in the same cell line transiently expressing the pGloSensor-22F protein following 16 h exposure to 1 μM PP2, <t>PP3,</t> eCF506 or an equal volume of DMSO. (A) Data are expressed as a percentage of maximum luminescence (% Max RLU) produced by DAMGO in DMSO exposed cells present in duplicate on each 96-well plate. The insert demonstrates c-Src phosphorylation at Y416 (upper panel) and total c-Src (lower panel) expression assessed using western blot in each cell lysate following overnight exposure and confirms inhibition of c-Src phosphorylation. (B) Data are expressed as a percentage of luminescence in each well measured prior to the addition of agonist (% control). Data from individual replicates were plotted and fitted with logistics functions to derive efficacy (E MAX ) and potency (EC 50 /IC 50 ) parameters, presented in . Data are the mean ± SEM of 5 replicates (A) and 8 replicates (B).
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    Concentration-response relationships of ( A ) DAMGO as a recruiter of β-arrestin2 to human μ receptors in PathHunter CHO cells and ( B ) DAMGO as an inhibitor of forskolin (30 μM) stimulated cAMP accumulation in the same cell line transiently expressing the pGloSensor-22F protein following 16 h exposure to 1 μM PP2, <t>PP3,</t> eCF506 or an equal volume of DMSO. (A) Data are expressed as a percentage of maximum luminescence (% Max RLU) produced by DAMGO in DMSO exposed cells present in duplicate on each 96-well plate. The insert demonstrates c-Src phosphorylation at Y416 (upper panel) and total c-Src (lower panel) expression assessed using western blot in each cell lysate following overnight exposure and confirms inhibition of c-Src phosphorylation. (B) Data are expressed as a percentage of luminescence in each well measured prior to the addition of agonist (% control). Data from individual replicates were plotted and fitted with logistics functions to derive efficacy (E MAX ) and potency (EC 50 /IC 50 ) parameters, presented in . Data are the mean ± SEM of 5 replicates (A) and 8 replicates (B).
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    Concentration-response relationships of ( A ) DAMGO as a recruiter of β-arrestin2 to human μ receptors in PathHunter CHO cells and ( B ) DAMGO as an inhibitor of forskolin (30 μM) stimulated cAMP accumulation in the same cell line transiently expressing the pGloSensor-22F protein following 16 h exposure to 1 μM PP2, <t>PP3,</t> eCF506 or an equal volume of DMSO. (A) Data are expressed as a percentage of maximum luminescence (% Max RLU) produced by DAMGO in DMSO exposed cells present in duplicate on each 96-well plate. The insert demonstrates c-Src phosphorylation at Y416 (upper panel) and total c-Src (lower panel) expression assessed using western blot in each cell lysate following overnight exposure and confirms inhibition of c-Src phosphorylation. (B) Data are expressed as a percentage of luminescence in each well measured prior to the addition of agonist (% control). Data from individual replicates were plotted and fitted with logistics functions to derive efficacy (E MAX ) and potency (EC 50 /IC 50 ) parameters, presented in . Data are the mean ± SEM of 5 replicates (A) and 8 replicates (B).
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    (A) The presence of either gain-of-function or loss-of-function mutations in cancer driver genes can lead to cancer (left) or rare Mendelian disorders (right) in different contexts. Most cancers result from somatic mutations that accumulate in a tissue-specific manner, whereas germline mutations are present in all cells of the body and cause a type of rare Mendelian disorder (e.g., neurodevelopmental disorder). (B) The HRAS Q61K mutation is an example of a known cancer mutation that drives different types of cancers that also causes Costello syndrome, a developmental disorder, when observed as a germline variant. (C) Workflow for extracting cancer mutations from Cancer Hotspots. Recurrent cancer mutations were filtered to 2,447 missense mutations. See main text for details. REVEL scores thresholds correspond to supporting evidence for pathogenicity <t>(PP3)</t> and for benign-ness <t>(BP4).</t> Created with Lucidchart. Created with BioRender.
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    Image Search Results


    Concentration-response relationships of ( A ) DAMGO as a recruiter of β-arrestin2 to human μ receptors in PathHunter CHO cells and ( B ) DAMGO as an inhibitor of forskolin (30 μM) stimulated cAMP accumulation in the same cell line transiently expressing the pGloSensor-22F protein following 16 h exposure to 1 μM PP2, PP3, eCF506 or an equal volume of DMSO. (A) Data are expressed as a percentage of maximum luminescence (% Max RLU) produced by DAMGO in DMSO exposed cells present in duplicate on each 96-well plate. The insert demonstrates c-Src phosphorylation at Y416 (upper panel) and total c-Src (lower panel) expression assessed using western blot in each cell lysate following overnight exposure and confirms inhibition of c-Src phosphorylation. (B) Data are expressed as a percentage of luminescence in each well measured prior to the addition of agonist (% control). Data from individual replicates were plotted and fitted with logistics functions to derive efficacy (E MAX ) and potency (EC 50 /IC 50 ) parameters, presented in . Data are the mean ± SEM of 5 replicates (A) and 8 replicates (B).

    Journal: bioRxiv

    Article Title: The c-Src inhibitor eCF506 diminishes opioid tolerance creating bias against β-arrestin2 recruitment

    doi: 10.1101/2025.07.01.662507

    Figure Lengend Snippet: Concentration-response relationships of ( A ) DAMGO as a recruiter of β-arrestin2 to human μ receptors in PathHunter CHO cells and ( B ) DAMGO as an inhibitor of forskolin (30 μM) stimulated cAMP accumulation in the same cell line transiently expressing the pGloSensor-22F protein following 16 h exposure to 1 μM PP2, PP3, eCF506 or an equal volume of DMSO. (A) Data are expressed as a percentage of maximum luminescence (% Max RLU) produced by DAMGO in DMSO exposed cells present in duplicate on each 96-well plate. The insert demonstrates c-Src phosphorylation at Y416 (upper panel) and total c-Src (lower panel) expression assessed using western blot in each cell lysate following overnight exposure and confirms inhibition of c-Src phosphorylation. (B) Data are expressed as a percentage of luminescence in each well measured prior to the addition of agonist (% control). Data from individual replicates were plotted and fitted with logistics functions to derive efficacy (E MAX ) and potency (EC 50 /IC 50 ) parameters, presented in . Data are the mean ± SEM of 5 replicates (A) and 8 replicates (B).

    Article Snippet: DAMGO ([D-Ala2, N-MePhe4, Gly-ol]-enkephalin) and forskolin (HelloBio, UK), morphine sulfate salt pentahydrate (#M8777), fentanyl citrate (#F3886), oxycodone hydrochloride (#BP1068; all from Merck, UK), TRV130 (#MBS3601671; CliniSciences, UK), buprenorphine hydrochloride (#2808), β-FNA (#0926), PP2 (#1407), PP3 (#2794), caffeic acid pYEEIE (#1935; all from Tocris, UK), DAS-5-oCRBN (#HY-163144; Cambridge Biosciences, UK) were prepared as 10 mM (DAMGO, morphine, fentanyl, buprenorphine, TRV130, PP2, PP3, DAS-5-oCRBN, caffeic acid-pYEEIE and eCF506), 20 mM (oxycodone), 25 mM (forskolin) or 100 mM (β-FNA) stock solutions in DMSO (DAMGO, forskolin, buprenorphine, β-FNA, PP2, PP3, DAS-5-oCRBN, caffeic acid-pYEEIE and eCF506) or water (fentanyl, morphine, oxycodone).

    Techniques: Concentration Assay, Expressing, Produced, Phospho-proteomics, Western Blot, Inhibition, Control

    Journal: bioRxiv

    Article Title: The c-Src inhibitor eCF506 diminishes opioid tolerance creating bias against β-arrestin2 recruitment

    doi: 10.1101/2025.07.01.662507

    Figure Lengend Snippet:

    Article Snippet: DAMGO ([D-Ala2, N-MePhe4, Gly-ol]-enkephalin) and forskolin (HelloBio, UK), morphine sulfate salt pentahydrate (#M8777), fentanyl citrate (#F3886), oxycodone hydrochloride (#BP1068; all from Merck, UK), TRV130 (#MBS3601671; CliniSciences, UK), buprenorphine hydrochloride (#2808), β-FNA (#0926), PP2 (#1407), PP3 (#2794), caffeic acid pYEEIE (#1935; all from Tocris, UK), DAS-5-oCRBN (#HY-163144; Cambridge Biosciences, UK) were prepared as 10 mM (DAMGO, morphine, fentanyl, buprenorphine, TRV130, PP2, PP3, DAS-5-oCRBN, caffeic acid-pYEEIE and eCF506), 20 mM (oxycodone), 25 mM (forskolin) or 100 mM (β-FNA) stock solutions in DMSO (DAMGO, forskolin, buprenorphine, β-FNA, PP2, PP3, DAS-5-oCRBN, caffeic acid-pYEEIE and eCF506) or water (fentanyl, morphine, oxycodone).

    Techniques: Inhibition, Concentration Assay, cAMP Assay

    Concentration-response relationships of DAMGO to recruit β-arrestin2 to human μ receptors in PathHunter CHO cells transiently overexpressing ( A ) full-length (residues 1-536; WT), ( B ) C-terminally truncated (residues 1-249) or ( C ) N-terminally truncated and catalytically inactive (residues 250-536(K298M)) human c-Src following 16-h exposure to PP2, PP3, eCF506 (300 nM) or an equal volume of DMSO. Data are expressed as a percentage of maximum luminescence (% Max RLU) produced by DAMGO in DMSO exposed cells present in duplicate on each plate i.e., cells overexpressing (A) WT, (B) Src 1-249 or (C) Src 250-536(K298M) are each expressed relative to the maximum β-arrestin2 recruitment of each construct caused by DAMGO. Data from individual replicates were plotted and fitted with logistics functions to derive efficacy (E MAX ) and potency (EC 50 ) parameters, presented in Supplementary Table 3. Data are the mean ± SEM of 5-6 replicates.

    Journal: bioRxiv

    Article Title: The c-Src inhibitor eCF506 diminishes opioid tolerance creating bias against β-arrestin2 recruitment

    doi: 10.1101/2025.07.01.662507

    Figure Lengend Snippet: Concentration-response relationships of DAMGO to recruit β-arrestin2 to human μ receptors in PathHunter CHO cells transiently overexpressing ( A ) full-length (residues 1-536; WT), ( B ) C-terminally truncated (residues 1-249) or ( C ) N-terminally truncated and catalytically inactive (residues 250-536(K298M)) human c-Src following 16-h exposure to PP2, PP3, eCF506 (300 nM) or an equal volume of DMSO. Data are expressed as a percentage of maximum luminescence (% Max RLU) produced by DAMGO in DMSO exposed cells present in duplicate on each plate i.e., cells overexpressing (A) WT, (B) Src 1-249 or (C) Src 250-536(K298M) are each expressed relative to the maximum β-arrestin2 recruitment of each construct caused by DAMGO. Data from individual replicates were plotted and fitted with logistics functions to derive efficacy (E MAX ) and potency (EC 50 ) parameters, presented in Supplementary Table 3. Data are the mean ± SEM of 5-6 replicates.

    Article Snippet: DAMGO ([D-Ala2, N-MePhe4, Gly-ol]-enkephalin) and forskolin (HelloBio, UK), morphine sulfate salt pentahydrate (#M8777), fentanyl citrate (#F3886), oxycodone hydrochloride (#BP1068; all from Merck, UK), TRV130 (#MBS3601671; CliniSciences, UK), buprenorphine hydrochloride (#2808), β-FNA (#0926), PP2 (#1407), PP3 (#2794), caffeic acid pYEEIE (#1935; all from Tocris, UK), DAS-5-oCRBN (#HY-163144; Cambridge Biosciences, UK) were prepared as 10 mM (DAMGO, morphine, fentanyl, buprenorphine, TRV130, PP2, PP3, DAS-5-oCRBN, caffeic acid-pYEEIE and eCF506), 20 mM (oxycodone), 25 mM (forskolin) or 100 mM (β-FNA) stock solutions in DMSO (DAMGO, forskolin, buprenorphine, β-FNA, PP2, PP3, DAS-5-oCRBN, caffeic acid-pYEEIE and eCF506) or water (fentanyl, morphine, oxycodone).

    Techniques: Concentration Assay, Produced, Construct

    (A) The presence of either gain-of-function or loss-of-function mutations in cancer driver genes can lead to cancer (left) or rare Mendelian disorders (right) in different contexts. Most cancers result from somatic mutations that accumulate in a tissue-specific manner, whereas germline mutations are present in all cells of the body and cause a type of rare Mendelian disorder (e.g., neurodevelopmental disorder). (B) The HRAS Q61K mutation is an example of a known cancer mutation that drives different types of cancers that also causes Costello syndrome, a developmental disorder, when observed as a germline variant. (C) Workflow for extracting cancer mutations from Cancer Hotspots. Recurrent cancer mutations were filtered to 2,447 missense mutations. See main text for details. REVEL scores thresholds correspond to supporting evidence for pathogenicity (PP3) and for benign-ness (BP4). Created with Lucidchart. Created with BioRender.

    Journal: PLOS Genetics

    Article Title: Leveraging cancer mutation data to inform the pathogenicity classification of germline missense variants

    doi: 10.1371/journal.pgen.1011540

    Figure Lengend Snippet: (A) The presence of either gain-of-function or loss-of-function mutations in cancer driver genes can lead to cancer (left) or rare Mendelian disorders (right) in different contexts. Most cancers result from somatic mutations that accumulate in a tissue-specific manner, whereas germline mutations are present in all cells of the body and cause a type of rare Mendelian disorder (e.g., neurodevelopmental disorder). (B) The HRAS Q61K mutation is an example of a known cancer mutation that drives different types of cancers that also causes Costello syndrome, a developmental disorder, when observed as a germline variant. (C) Workflow for extracting cancer mutations from Cancer Hotspots. Recurrent cancer mutations were filtered to 2,447 missense mutations. See main text for details. REVEL scores thresholds correspond to supporting evidence for pathogenicity (PP3) and for benign-ness (BP4). Created with Lucidchart. Created with BioRender.

    Article Snippet: We grouped these CH mutations by REVEL scores using the ClinGen-proposed PP3/BP4 score thresholds ( ) [ ].

    Techniques: Mutagenesis, Variant Assay