0.12ug of CSNK1a1 recombinant human protein (ThermoFisher Scientific) and 0.2ug of PRMT1 recombinant human protein (SignalChem) were combined in 25uL of kinase assay buffer (50 mM Tris-HCl, 10 mM MgCl2, 0.1 mM EDTA, 2 mM DTT, 0.01% Brij 35, 200 μM ATP, pH 7.5), and incubated for 60min at 30°C. The entire reactions were heat inactivated and separated on a SDS-PAGE gel. Anti-phospho-Serine antibody (ThermoFisher Scientific) were used at 2ug/mL to detect serine phosphorylation. Since both recombinant proteins have GST-tag, anti-GST (Cell Signaling 1:2000) were used to detect input.
Anti phospho serine antibody
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Anti-phospho-Serine antibody is a laboratory reagent used to detect and quantify the presence of phosphorylated serine residues in proteins. It is a useful tool for studying protein phosphorylation, a common post-translational modification that plays a crucial role in various cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Anti gst, by Cell Signaling Technology (1 mentions)
Anti acetylated lysine, by Cell Signaling Technology (1 mentions)
Anti peroxiredoxin 2, by Abcam (1 mentions)
Ipg buffer ph 4 7, by GE Healthcare (1 mentions)
Horseradish peroxidase conjugated secondary antibody, by Enzo Life Sciences (1 mentions)
Anti phospho threonine, by Cell Signaling Technology (1 mentions)
Mg132, by Merck Group (1 mentions)
Protein g sepharose, by GE Healthcare (1 mentions)
Hydrogen peroxide, by Merck Group (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Anti p47phox antibody, by Santa Cruz Biotechnology (1 mentions)
Dynabeads protein g immunoprecipitation kit, by Thermo Fisher Scientific (1 mentions)
Cell lysis buffer, by Cell Signaling Technology (1 mentions)
3 protocols using anti phospho serine antibody
1
In Vitro Kinase Assay of CSNK1a1 and PRMT1
2
Antibody Reagents for Peroxiredoxin Analysis
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Anti-peroxiredoxin-1, anti-peroxiredoxin-2, anti-peroxiredoxin-3, and anti-peroxiredoxin-SO3 were obtained from Abcam (Cambridge, MA, USA). Anti-peroxiredoxin-6 and β-actin were purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Anti-phosphothreonine and anti-acetylated lysine were purchased from Cell Signaling Technology (Danvers, MA, USA). Anti-phosphoserine antibody was obtained from Thermo Fisher Scientific (Waltham, MA, USA). Horseradish peroxidase-conjugated secondary antibodies were purchased from Enzo Life Sciences (Farmingdale, NY, USA). Protein G-Sepharose and IPG buffer pH 4–7 were obtained from GE Healthcare Life Sciences (Pittsburgh, PA, USA). Hydrogen peroxide, MG132, and cycloheximide were purchased from Sigma Aldrich (St. Louis, MO, USA). Lactacystin was obtained from AG Scientific, Inc. (San Diego, CA, USA).
3
Immunoprecipitation and Immunoblotting of p47phox
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BMDM were lysed in cell lysis buffer (9803, Cell Signaling Technology, Danvers, MA, USA) with protease inhibitor cocktails and PMSF (P8340 and 93482, Sigma-Aldrich). The lysed cell protein was immunoprecipitated with anti-p47phox antibody (sc-14015, Santa Cruz Biotechnology) using Dynabeads protein G immunoprecipitation kit (10007D, Thermo Fisher Scientific). The immunoprecipitated proteins were then subjected to immunoblotting analysis using anti-phosphoserine antibody (61–8100, Thermo Fisher Scientific) and anti-gp91phox antibody (sc-5827, Santa Cruz Biotechnology), respectively.
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