Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line untreated (-) or treated (+) with 200 nM PMA for 20 minutes. PVDF Membrane was probed with 1 µg/mL of Human Phospho-GSK-3 beta (S9) Monoclonal Antibody (Catalog # MAB25062) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-GSK-3 beta (S9) at approximately 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Phospho-GSK‑3 beta (S9) in HeLa Human Cell Line.

GSK-3 beta phosphorylated at S9 was detected in immersion fixed HeLa human cervical epithelial carcinoma cells, unstimulated (lower panel) or stimulated (upper panel) with PMA, using Mouse Anti-Human Phospho-GSK-3 beta (S9) Monoclonal Antibody (Catalog # MAB25062) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Phospho-GSK-3 beta (S9) in HeLa Human Cell Line by Flow Cytometry.

HeLa human cervical epithelial carcinoma cell line untreated (open histogram) or treated with 120 ng/mL PMA for 15 minutes (filled histogram) was stained with Human Phospho-GSK-3 beta (S9) Monoclonal Antibody (Catalog # MAB25062) or isotype control antibody (Catalog # MAB002, not shown), followed by Fluorescein-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0103B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

Applications for Human Phospho-GSK-3 beta (S9) Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell line stimulated with PMA

Intracellular Staining by Flow Cytometry

2.5 µg/10⁶ cells
Sample: HeLa human cervical epithelial carcinoma cell line treated with PMA, fixed with paraformaldehyde and permeabilized with saponin

Western Blot

1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line treated with PMA

Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 4 using MAB25062 in the following applications:

Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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 beta

Glycogen Synthase Kinase-3 (GSK-3) is a serine/threonine kinase initially identified asan inhibitor of glycogen synthase. Two isoforms (GSK-3 alpha and GSK-3 beta) share 85% amino acid identity. GSK-3 beta, inhibited by phosphorylation at S9 by Akt, is involved in energy metabolism, body pattern formation, and neuronal cell development.

Long Name

Glycogen Synthase Kinase 3 beta

Alternate Names

GSK3 beta, GSK3B

Entrez Gene IDs

2932 (Human); 56637 (Mouse); 84027 (Rat)

Gene Symbol

GSK3B

Additional GSK-3 beta Products

Product Documents for Human Phospho-GSK-3 beta (S9) Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human Phospho-GSK-3 beta (S9) Antibody

For research use only

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Reviews

Calculators
Flow Cytometry

Human Phospho-GSK-3 beta (S9) Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB25062

Key Product Details

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Applications

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Antibody Source

View all GSK-3 beta Antibodies »

Product Specifications

Immunogen

Specificity

Clonality

Host

Isotype

Scientific Data Images for Human Phospho-GSK-3 beta (S9) Antibody

Detection of Human Phospho-GSK‑3 beta (S9) by Western Blot.