Human/Mouse/Rat GSK‑3 alpha / beta Antibody

R&D Systems | Catalog # AF2157

R&D Systems
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Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Human, Mouse, Rat

Applications

Validated:

Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunoprecipitation

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG
View all GSK-3 alpha/beta Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human GSK-3 beta
Accession # P49841

Specificity

Detects human, mouse, and rat GSK-3 beta and the closely related GSK-3 alpha.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human/Mouse/Rat GSK‑3 alpha / beta Antibody

Detection of Human GSK-3a/ beta antibody by Western Blot.

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-3a/ 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-3a/ beta at approximately 46 and 51 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
GSK-3a/ beta antibody in HeLa Human Cell Line by Immunocytochemistry (ICC).

GSK‑3 alpha / beta in HeLa Human Cell Line.

GSK-3a/ beta was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Rabbit Anti-Human/Mouse/Rat GSK-3a/ 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-3a/ beta antibody in HeLa Human Cell Line antibody by Flow Cytometry.

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-3a/ 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.
Detection of Mouse GSK-3 alpha/beta by Western Blot

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

GSK3 binds to scaffold formed by histidine-rich cytoplasmic loop domains within the ZIP6-ZIP10 heteromer.(a) Domain organization of ZIP10 and ZIP6 and deletion constructs used in this work to test the hypothesis that a cytoplasmic loop connecting transmembrane (TM) domains III and IV is essential for binding to GSK3 binding. (b) Control Western blot documenting ZIP6 expression products in eluates shown in panel ‘e’. (c) The co-expression of wild-type ZIP6 and ZIP10 proteins comprising intact cytoplasmic loop domains connecting TM domains III and IV is essential for binding of GSK3. GSK3-directed Western blot analysis of cellular lysates and co-immunoprecipitation eluates from in vivo formaldehyde crosslinked cells expressing different combinations of wild-type and/or mutant ZIP6/ZIP10. Note that in addition to protein bands reflecting the expected monomeric molecular weights of GSK3 and ZIP6, the Western blots in panels ‘d’ and ‘e’ revealed the appearance of high molecular weight crosslinked bands, whose appearance depended on the application of the in vivo formaldehyde crosslinking step. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/28098160), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse/Rat GSK‑3 alpha / beta 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

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

Intracellular Staining by Flow Cytometry

2.5 µg/106 cells
Sample: HeLa Human cell line fixed with paraformaldehyde and permeabilized with saponin

Western Blot

0.1 µg/mL
Sample: 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

Reviewed Applications

Read 4 reviews rated 4.5 using AF2157 in the following applications:

Flow Cytometry Panel Builder

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen 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. *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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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.

Long Name

Glycogen Synthase Kinase 3

Alternate Names

DKFZp686D0638, EC 2.7.11, EC 2.7.11.26, glycogen synthase kinase 3 alpha, glycogen synthase kinase-3 alpha, GSK-3 alpha

UniProt

P49841

Additional GSK-3 alpha/beta Products

Product Documents for Human/Mouse/Rat GSK‑3 alpha / beta Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human/Mouse/Rat GSK‑3 alpha / beta Antibody

For research use only

Citations for Human/Mouse/Rat GSK‑3 alpha / beta Antibody

Customer Reviews for Human/Mouse/Rat GSK‑3 alpha / beta Antibody (4)

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4 Customer Ratings
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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • Human/Mouse/Rat GSK-3 alpha / beta Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Lung single-cell suspension
    Species: Mouse
    Verified Customer | Posted 06/06/2019
    Human/Mouse/Rat GSK‑3 alpha / beta Antibody AF2157
  • Human/Mouse/Rat GSK-3 alpha / beta Antibody
    Name: Kurt Patterson
    Application: Western Blot
    Sample Tested: Colon cancer cell line
    Species: Human
    Verified Customer | Posted 09/14/2018
    Human/Mouse/Rat GSK‑3 alpha / beta Antibody AF2157
  • Human/Mouse/Rat GSK-3 alpha / beta Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: IPS2 induced pluripotent stem cells
    Species: Mouse
    Verified Customer | Posted 01/25/2018
    Human/Mouse/Rat GSK‑3 alpha / beta Antibody AF2157
  • Human/Mouse/Rat GSK-3 alpha / beta Antibody
    Name: Dharmaraja Allimuthu
    Application: Western Blot
    Sample Tested: Pancreatic cancer cells
    Species: Human
    Verified Customer | Posted 12/27/2017
    Human/Mouse/Rat GSK‑3 alpha / beta Antibody AF2157

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