Detection of Human GSK‑3 alpha / beta by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, HT‑29 human colon adenocarcinoma cell line, MDA‑MB‑468 and MCF‑7 human breast cancer cell line. PVDF membrane was probed with 0.1 µg/mL of Rabbit Anti-Human/Mouse/ Rat GSK‑3 alpha / beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2157) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). Specific bands were detected for GSK‑3 alpha / beta at approximately 46 and 51 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
GSK‑3 alpha / beta was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Rabbit Anti-Human/Mouse/Rat GSK‑3 alpha / beta Antigen Affinity-purified Polyclonal Antibody (Catalog # AF2157) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; Catalog # NL004) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Detection of GSK‑3 alpha / beta in HeLa Human Cell Line by Flow Cytometry.
HeLa human cervical epithelial carcinoma cell line was stained with Rabbit Anti-Human/Mouse/Rat GSK‑3 alpha / beta Antigen Affinity‑purified Polyclonal Antibody (Catalog # AF2157, filled histogram) or control antibody (Catalog # AB‑105‑C, open histogram), followed by Allophycocyanin-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # F0111). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: GSK-3 alpha/beta
GSK-3 is a Ser/Thr kinase first identified as an inactivator of Glycogen Synthase. GSK-3 acts as a multifunctional downstream switch that determines the output of numerous signaling pathways. There are two mammalian GSK-3 isoforms encoded by distinct genes, GSK-3 alpha and GSK-3 beta, which are structurally similar, but functionally non-identical. GSK-3a is inhibited by phosphorylation at S21 by Akt and other kinases. GSK-3 alpha and GSK-3 beta share 85% amino acid identity. Dysregulated GSK-3 has been implicated in several diseases including type II diabetes, Alzheimer's disease, bipolar disorder, and cancer.
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