Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-GSK3 alpha antibody [EP793Y] (ab40870)

Submit a review Submit a question References (6)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP793Y] to GSK3 alpha
  • Suitable for: ICC/IF, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-GSK3 alpha antibody [EP793Y]
    See all GSK3 alpha primary antibodies

  • Description

    Rabbit monoclonal [EP793Y] to GSK3 alpha

  • Host species

    Rabbit

  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Pmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human GSK3 alpha aa 50-150 (N terminal). The exact sequence is proprietary.
    Database link: P49840

  • Positive control

    • HeLa cell lysate; human breast carcinoma tissue. ICC/IF: HeLa cells

  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Applications

Our Abpromise guarantee covers the use of ab40870 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
ICC/IF 1/200.
WB 1/5000. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

Images

  • Western blot - Anti-GSK3 alpha antibody [EP793Y] (ab40870)
    Western blot - Anti-GSK3 alpha antibody [EP793Y] (ab40870)
    All lanes : Anti-GSK3 alpha antibody [EP793Y] (ab40870) at 1/1000 dilution

    Lane 1 : Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 2 : GSK3A knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate
    Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 4 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 51 kDa
    Observed band size: 51 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab40870 observed at 51 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab40870 Anti-GSK3 alpha antibody [EP793Y] was shown to specifically react with GSK3 alpha in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266742 (knockout cell lysate ab257979) was used. Wild-type and GSK3 alpha knockout samples were subjected to SDS-PAGE. ab40870 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-GSK3 alpha antibody [EP793Y] (ab40870)
    Western blot - Anti-GSK3 alpha antibody [EP793Y] (ab40870)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: GSK3 alpha knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: HepG2 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab40870 observed at 51 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab40870 was shown to specifically react with GSK3 alpha in wild-type HAP1 cells. No band was observed when GSK3 alpha knockout samples were examined. Wild-type and GSK3 alpha knockout samples were subjected to SDS-PAGE. ab40870 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/5000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-GSK3 alpha antibody [EP793Y] (ab40870)
    Immunocytochemistry/ Immunofluorescence - Anti-GSK3 alpha antibody [EP793Y] (ab40870)

    Immunocytochemistry/ Immunofluorescence analysis of HeLa (human cervix adenocarcinoma epithelial cell) cells labeling GSK3 alpha with purified ab40870 at 1/200 dilution (10 µg/mL). Cells were fixed in 100% Methanol. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 alpha antibody [EP793Y] (ab40870)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 alpha antibody [EP793Y] (ab40870)

    ab4870 at 1/50 dilution, staining GSK3 alpha in Human breast carcinoma tissue by Immunohistochemistry, Paraffin embedded section.

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Western blot - Anti-GSK3 alpha antibody [EP793Y] (ab40870)
    Western blot - Anti-GSK3 alpha antibody [EP793Y] (ab40870)
    Anti-GSK3 alpha antibody [EP793Y] (ab40870) at 1/5000 dilution + 10ug HeLa cell lysate.

    Predicted band size: 51 kDa
    Observed band size: 51 kDa

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (6)

Publishing research using ab40870? Please let us know so that we can cite the reference in this datasheet.

ab40870 has been referenced in 6 publications.

  • Duthie A  et al. Recruitment, Retainment, and Biomarkers of Response; A Pilot Trial of Lithium in Humans With Mild Cognitive Impairment. Front Mol Neurosci 12:163 (2019). PubMed: 31316348
  • Sang B  et al. Ras-AKT signaling represses the phosphorylation of histone H1.5 at threonine 10 via GSK3 to promote the progression of glioma. Artif Cells Nanomed Biotechnol 47:2882-2890 (2019). PubMed: 31307224
  • Krishnankutty A  et al. In vivo regulation of glycogen synthase kinase 3ß activity in neurons and brains. Sci Rep 7:8602 (2017). PubMed: 28819213
  • Shukla S  et al. Cucurbitacin B inhibits the stemness and metastatic abilities of NSCLC via downregulation of canonical Wnt/ß-catenin signaling axis. Sci Rep 6:21860 (2016). WB . PubMed: 26905250
  • Yamaguchi N  et al. Dysfunctional ryanodine receptor and cardiac hypertrophy: role of signaling molecules. Am J Physiol Heart Circ Physiol 300:H2187-95 (2011). WB ; Mouse . PubMed: 21421818
  • Yamauchi K & Kurosaka A Expression and function of glycogen synthase kinase-3 in human hair follicles. Arch Dermatol Res 302:263-70 (2010). PubMed: 19705135

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

There are currently no Customer reviews or Questions for ab40870.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com