|GSK‑3 alpha in Human Thymus. GSK‑3 alpha was detected in immersion fixed paraffin-embedded sections of human thymus using 5 µg/mL Human/Mouse/Rat Phospho-GSK‑3 alpha (S21) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4125) overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained with the Anti-Rabbit HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS005) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.|
|Detection of Human and Mouse Phospho-GSK‑3 alpha (S21) by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 200 nM PMA for 20 minutes and MCF‑7 human breast cancer cell line untreated or treated with 100 ng/mL Recombinant Human IGF-1 (Catalog # 291-G1) for 15 minutes and NIH‑3T3 mouse embryonic fibroblast cell line untreated or treated with 10 ng/mL PMA for 20 minutes. PVDF membrane was probed with 1 µg/mL of Human/Mouse/Rat Phospho-GSK‑3 alpha (S21) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF4125), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band was detected for Phospho-GSK‑3 alpha (S21) at approximately 51 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
Glycogen Synthase Kinase-3 (GSK-3) is a serine/threonine kinase initially identified as an inhibitor of glycogen synthase. Two isoforms (GSK-3 alpha and GSK-3 beta ) share 85% amino acid identity. GSK-3 alpha is inhibited by phosphorylation at S21 by Akt and other kinases. Deregulated GSK-3 has been implicated in several diseases including type II diabetes, Alzheimer’s disease, bipolar disorder, and cancer.
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