Human/Mouse/Rat Phospho-GSK-3 alpha/ beta (S21/S9) Antibody

Name: Human/Mouse/Rat Phospho-GSK-3 alpha/ beta (S21/S9) Antibody
Brand: R&D Systems
SKU: AF1590
Rating: 5 (1 reviews)

R&D Systems, part of Bio-Techne | Catalog # AF1590

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Citations (16)

Product Datasheet / COA / SDS

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Product Specifications
Scientific Data
Applications
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Immunocytochemistry, Western Blot

Cited:

Flow Cytometry, Immunohistochemistry, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

View all GSK-3 alpha/beta Antibodies »

Product Specifications

Immunogen

Phosphopeptide containing GSK-3 beta S9 site

Specificity

Detects human, mouse, and rat GSK-3 beta when phosphorylated at S9, and human, mouse, and rat GSK‑3 alpha when phosphorylated at S21.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat Phospho-GSK-3 alpha/ beta (S21/S9) Antibody

Detection of Human Phospho-GSK‑3 alpha (S21) and GSK-3 beta (S9) 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 (291-G1) for 15 minutes. PVDF membrane was probed with 0.2 µg/mL of Human/Mouse/Rat Phospho-GSK-3a/ beta (S21/S9) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1590), followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). Specific bands were detected for Phospho-GSK-3a (S21) and GSK-3 beta (S9) at approximately 51 and 46 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Phospho-GSK-3 alpha/ beta (S21/S9) in HeLa Human Cell Line.

Phospho-GSK-3 alpha/ beta (S21/S9) was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line treated with PMA (left panel; positive staining) and untreated HeLa cell line (right panel; negative control) using Rabbit Anti-Human/Mouse/Rat Phospho-GSK-3 alpha/ beta (S21/S9) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1590) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red; NL004) and counterstained with DAPI (blue). Specific staining was localized to nuclei. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of Rat GSK-3 alpha/beta by Western Blot

HYS-32 modulates GSK3 beta phosphorylation.(A) Cell lysates from control astrocytes (Con) or astrocytes treated for 2, 6, 12, or 24 h with 5 μM HYS-32 were subjected to 10% SDS-PAGE, and analyzed by immunoblotting with antibodies against GSK3 beta-pTyr216, GSK3 beta-pSer9, or GAPDH. (B) Densitometric analyses of GSK3 beta-pTyr216 and GSK3 beta-pSer9 expressed as the density of the bands in the treated groups relative to the control. *p<0.05 compared to control using one-way ANOVA with Dunnett’s post-hoc test. The results were collected from five independent experiments. (C) According to the data in (B), the stacked bar graph showing the relative percentage of phospho-GSK3 beta at indicated time. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0126217), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Applications for Human/Mouse/Rat Phospho-GSK-3 alpha/ beta (S21/S9) Antibody

Application

Recommended Usage

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed HeLa human cervical epithelial carcinoma cell treated with PMA

Western Blot

0.2 µg/mL
Sample: PMA-treated HeLa human cervical epithelial carcinoma cell line and MCF-7 human breast cancer cell line treated with Recombinant Human IGF-1 (Catalog # 291-G1)

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

Reviewed Applications

Read 1 review rated 5 using AF1590 in the following applications:

Western Blot (1 Review)

Formulation, Preparation, and Storage

Purification

Antigen and protein A Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*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 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.