Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)

Overview

  • Product nameAnti-GSK3 beta (phospho S9) antibody [EPR2286Y]
    See all GSK3 beta primary antibodies
  • Description
    Rabbit monoclonal [EPR2286Y] to GSK3 beta (phospho S9)
  • Specificityab75814 only detects GSK3 beta phosphorylated on Serine 9.
  • Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity
    Reacts with: Human
  • Immunogen

    A phospho-specific peptide corresponding to residues surrounding Serine 9 of human GSK3 beta

  • Positive control
    • 293T cell lysates, untreated or treated with Calyculin A and human ovarian carcinoma tissue.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Applications

Our Abpromise guarantee covers the use of ab75814 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
WB 1/10000 - 1/20000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF 1/100 - 1/250.
  • Application notesIs unsuitable for Flow Cyt or IP.
  • 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 (phospho S9) antibody [EPR2286Y] images

    • Cell line: HeLa (human cervix adenocarcinoma)

      Target: Ab75814 anti-GSK3 beta (phospho S9)

      Secondary: ab150077 AlexaFluor®488 Goat anti-Rabbit 

      Fixative: 4% PFA

      Permeabilization: 0.1% Triton-X

      Nuclear counter stain: DAPI

      Confocal image showed increased cytoplasmic staining after Calyculin A (CA) (ab141784 100ng/ml, 10min) treatment on Hela cells. The LP treatment decreased the increased cytoplasmic staining caused by CA.

      Ab32391 was used as a control for ab75814. The results showed cytoplasmic staining on untreated, CA and CA+LP treated Hela cells.

    • All lanes : Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814) at 1/20000 dilution

      Lane 1 : Untreated HeLa cell lysate
      Lane 2 : HeLa cells were starved for 3 hours then treated with Calyculin A (100nM) for 30 minutes.
      Lane 3 : HeLa cells were starved for 3 hours then treated with Calyculin A (100nM) for 30 minutes. The membrane was treated with alkaline phosphatase.

      Lysates/proteins at 10 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat anti-rabbit IgG H&L (HRP))

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


      Exposure time : 5 seconds

      Blocking buffer: 5% NFDM/TBST for the first 3 lanes, 2% BSA/TBST for the second 3 lanes.

      Dilution buffer: 5% NFDM/TBST for the first 3 lanes, 2% BSA/TBST for the second 3 lanes.

    • Immunohistochemical analysis of paraffin-embedded human breast tissue labelled with untreated ab75814 (phospho) (top-left) at a dilution of 1/1000, alkaline phosphatase treated ab75814 (phospho) (top-right) at a dilution of 1/1000, untreated ab32391 (bottom-left) at a dilution of 1/1000 and alkaline phophatase treated ab32391 (bottom-right) at a dilution of 1/1000. Ab97051 was used as secondary antibody at a dilution of 1/500 and counterstained with hematoxylin.

    • Dot blot analysis of GSK3 beta (pS9) peptide (Lane 1) and GSK3 beta non-phospho peptide (Lane 2) labelling GSK3 (pS9) with purified ab75814 at a dilution of 1/1000. ab97051 (Peroxidase conjugated goat anti-rabbit IgG (H+L)) was used as the secondary antibody at a dilution of 1/100000.

      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure time: 3 minutes.

    • Immunohistochemical analysis of paraffin-embedded human breast cancer tissue labelled with untreated ab75814 (top-left) at a dilution of 1/1000, alkaline phosphatase treated ab75814 (top-right) at a dilution of 1/1000, untreated ab32391 (bottom-left) at a dilution of 1/1000 and alkaline phophatase treated ab32391 (bottom-right) at a dilution of 1/1000. Ab97051 was used as secondary antibody at a dilution of 1/500 and counterstained with hematoxylin.

    • All lanes : Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814) at 1/20000 dilution

      Lane 1 : 293T cell lysates, untreated
      Lane 2 : 293T cell lysates, treated with Calyculin A

      Lysates/proteins at 10 µg per lane.

      Secondary
      goat anti-rabbit HRP at 1/2000 dilution

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

    References for Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)

    This product has been referenced in:
    • Sun J  et al. Astragaloside IV protects new born rats from anesthesia-induced apoptosis in the developing brain. Exp Ther Med 12:1829-1835 (2016). Read more (PubMed: 27588101) »
    • Chen YC  et al. High mobility group box 1-induced epithelial mesenchymal transition in human airway epithelial cells. Sci Rep 6:18815 (2016). WB ; Human . Read more (PubMed: 26739898) »

    See all 11 Publications for this product

    Product Wall

    Application Western blot
    Loading amount 150 µg
    Gel Running Conditions Reduced Denaturing (gel 10% SDS-PAGE)
    Sample Mouse Tissue lysate - whole (Brain)
    Specification Brain
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Dr. Karine Thibault

    Verified customer

    Submitted Nov 10 2014

    Thank you for contacting us.

    Both of these antibodies are equally suitable for detecting the phospho form of GSK3 beta. Please note ab75814 is a rabbit monoclonal antibody.

    I hope this information is helpful to you. Please do not hesi...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (293FT cells)
    Loading amount 40 µg
    Specification 293FT cells
    Treatment Transfection
    Gel Running Conditions Non-reduced Denaturing (10%)
    Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Username

    Chien Hsin Lee

    Verified customer

    Submitted Sep 09 2009

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