Phospho gsk 3α β ser21 9 antibody

Manufactured by Cell Signaling Technology

The Phospho-GSK-3α/β (Ser21/9) Antibody is a primary antibody used to detect the phosphorylation of glycogen synthase kinase-3 alpha and beta at serine 21 and 9 residues, respectively. This antibody can be used in various immunoassay techniques to study the activation state of these kinases.

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Lab products found in correlation

P62 antibody, by Cell Signaling Technology (1 mentions) Cleaved caspase 3 antibody, by Cell Signaling Technology (1 mentions) Gapdh rabbit mab, by Cell Signaling Technology (1 mentions) Nlrp3 rabbit mab, by Cell Signaling Technology (1 mentions) Mtor rabbit mab, by Cell Signaling Technology (1 mentions) Protease inhibitor, by Merck Group (1 mentions) Pvdf membrane, by Bio-Rad (1 mentions) Ripa lysis buffer, by Beyotime (1 mentions) Bca kit, by Beyotime (1 mentions) Supersignal west pico ecl, by Thermo Fisher Scientific (1 mentions) Pka c α antibody, by Cell Signaling Technology (1 mentions)

2 protocols using phospho gsk 3α β ser21 9 antibody

Western Blot Analysis of Protein Signaling

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Protein lysates were extracted from brain tissues or culture cells by RIPA lysis buffer (Beyotime, China) supplemented with protease inhibitors (Sigma, USA). Protein concentration was measured by BCA kit (Beyotime, China). A total of 20 μg protein was separated by 10% SDS-PAGE and transferred onto PVDF membranes (Biorad, USA). Then, the membranes were blocked by 5% fat-free milk for 1 h at room temperature, incubated with first antibodies at 4 °C overnight and corresponding second antibodies for 1 h at room temperature. Then antibodies used in our study were: iNOS Rabbit mAb (CST#13120, 1: 1000), Arginase-1 Rabbit mAb (CST#93668, 1: 1000), GAPDH Rabbit mAb (CST#2118, 1: 1000), LC3A/B Rabbit mAb (CST#12741, 1: 1000), p62 Antibody (CST#5114, 1: 1000), Phospho-Akt (Ser473) Antibody (CST#9271, 1: 1000), Akt Antibody (CST#9272, 1: 1000), Phospho-mTOR (Ser2448) Antibody (CST#2971, 1: 1000), mTOR Rabbit mAb (CST#2983, 1: 1000), pro-IL-1β Rabbit mAb (CST#31202, 1: 1000), Cleaved-IL-1β Rabbit mAb (CST#63124, 1: 1000), pro-Caspase-3 Antibody (CST#9662, 1: 1000), Cleaved Caspase-3 Antibody (CST#9661, 1: 1000), Phospho-GSK-3α/β (Ser21/9) Antibody (CST#9331, 1: 1000), GSK-3α/β Rabbit mAb (CST#5676, 1: 1000) and NLRP3 Rabbit mAb (CST#15101, 1: 1000) were all obtained from Cell Signaling Technology (USA).

Kang C., Sang Q., Liu D., Wang L., Li J, & Liu X. (2024). Polyphyllin I alleviates neuroinflammation after cerebral ischemia–reperfusion injury via facilitating autophagy-mediated M2 microglial polarization. Molecular Medicine, 30, 59.

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Immunoblotting of Phosphoproteins

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The protein fractions were separated on SDS-PAGE and transferred to PVDF membrane by Bio-Rad Laboratories, Hercules, CA wet-transfer apparatus. Nonspecific binding sites were blocked with 5% skimmed milk. The PVDF paper was then incubated with primary antibody: rabbit polyclonal Anti-PDE4 antibody (Abcam, Cambridge, MA #ab14628), rabbit polyclonal phospho-GSK-3α/β (Ser21/9) antibody (Cell Signaling, Danvers, MA #9331), anti-GSK3 α/β mouse monoclonal antibody (44610; Invitrogen (Thermo Fisher Scientific), Waltham, MA), rabbit polyclonal PKA C-α antibody (Cell Signaling #4782), rabbit polyclonal Phospho-PP1α (Thr320) antibody (Cell Signaling #2581), anti-PP1gamma2 (PPP1CC2) antibody (commercially prepared using a synthetic peptide corresponding to the 22 amino acids at the carboxy terminus of PPP1CC2 as the antigen); all the primary antibodies were used at 1:1000 dilution. This was followed by horseradish peroxidase-conjugated secondary IgG (1:2500 dilution). Immunoreactive bands were visualized using a chemiluminescent substrate (Thermo Scientific Super Signal West Pico ECL). The PVDF membrane was then reprobed with either rabbit polyclonal β-tubulin (for whole cell extracts) or mouse monoclonal β-actin (for soluble protein extracts) antibody to verify the uniformity of the sample loading.

Dey S., Goswami S., Eisa A., Bhattacharjee R., Brothag C., Kline D, & Vijayaraghavan S. (2018). Cyclic AMP and glycogen synthase kinase 3 form a regulatory loop in spermatozoa. Journal of cellular physiology, 233(9), 7239-7252.

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