Overview

  • Product nameAnti-GSK3 beta antibody
    See all GSK3 beta primary antibodies
  • Description
    Rabbit polyclonal to GSK3 beta
  • Tested applicationsSuitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Cow, PigDoes not react with: Mouse, Rat
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human GSK3 beta.

    (Peptide available as ab73172.)

  • Positive control
    • This antibody gave a positive signal in the following Human Whole Cell lysates: Jurkat - Staurosporine Treated (24hr, 500nM) and HeLa

Associated products

Applications

Our Abpromise guarantee covers the use of ab73173 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 Use a concentration of 5 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Target

  • FunctionParticipates in the Wnt signaling pathway. Implicated in the hormonal control of several regulatory proteins including glycogen synthase, MYB and the transcription factor JUN. Phosphorylates JUN at sites proximal to its DNA-binding domain, thereby reducing its affinity for DNA. Phosphorylates MUC1 in breast cancer cells, and decreases the interaction of MUC1 with CTNNB1/beta-catenin. Phosphorylates CTNNB1/beta-catenin. Phosphorylates SNAI1. Plays an important role in ERBB2-dependent stabilization of microtubules at the cell cortex. Prevents the phosphorylation of APC and CLASP2, allowing its association with the cell membrane. In turn, membrane-bound APC allows the localization of MACF1 to the cell membrane, which is required for microtubule capture and stabilization. Phosphorylates MACF1 and this phosphorylation inhibits the binding of MACF1 to microtubules which is critical for its role in bulge stem cell migration and skin wound repair.
  • Tissue specificityExpressed in testis, thymus, prostate and ovary and weakly expressed in lung, brain and kidney.
  • Sequence similaritiesBelongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. GSK-3 subfamily.
    Contains 1 protein kinase domain.
  • Post-translational
    modifications
    Phosphorylated by AKT1 and ILK1. Activated by phosphorylation at Tyr-216.
  • Cellular localizationCytoplasm. Nucleus. Cell membrane. The phosphorylated form shows localization to cytoplasm and cell membrane. The MEMO1-RHOA-DIAPH1 signaling pathway controls localization of the phosophorylated form to the cell membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Glycogen Synthase Kinase 3 Beta antibody
    • Glycogen synthase kinase-3 beta antibody
    • GSK 3 beta antibody
    • GSK-3 beta antibody
    • GSK3B antibody
    • GSK3B_HUMAN antibody
    • GSK3beta isoform antibody
    • Serine/threonine-protein kinase GSK3B antibody
    see all

Anti-GSK3 beta antibody images



  • Predicted band size : 47 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: GSK3 beta knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: A431 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab73173 observed at 47 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab73173 was shown to recognize GSK3 beta when GSK3 beta knockout samples were used, along with additional cross-reactive bands. Wild-type and GSK3 beta knockout samples were subjected to SDS-PAGE. ab73173 and ab8245 (loading control to GAPDH) were diluted at 1 µg/ml and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • All lanes : Anti-GSK3 beta antibody (ab73173) at 1 µg/ml

    Lane 1 : Jurkat Whole Cell Lysate - Staurosporine Treated (24hr, 500nM)
    Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 47 kDa
    Observed band size : 47 kDa


    Exposure time : 20 minutes
  • ICC/IF image of ab73173 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab73173, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) HeLa cells at 5µg/ml.
  • IHC image of GSK3 beta staining in human testis formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab71373, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • ICC/IF image of ab73173 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab73173, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 47 kDa


    Exposure time : 90 seconds
    ab60863 is a tagged recombinant protein with an expected molecular weight of 73kDa.

References for Anti-GSK3 beta antibody (ab73173)

ab73173 has not yet been referenced specifically in any publications.

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (Testis)
Specification Testis
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Feb 14 2013

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"