Recombinant
RabMAb

Recombinant Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] - BSA and Azide free (ab219359)

Overview

  • Product name

    Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] - BSA and Azide free
    See all GSK3 beta primary antibodies
  • Description

    Rabbit monoclonal [EPR2286Y] to GSK3 beta (phospho S9) - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, ICC/IF, WB, Dot blotmore details
    Unsuitable for: Flow Cyt or IP
  • Species reactivity

    Reacts with: Human
  • Positive control

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

    Ab219359 is the carrier-free version of ab75814. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab219359 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.??

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

    This product is a recombinant rabbit monoclonal antibody.

Applications

Our Abpromise guarantee covers the use of ab219359 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
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
Dot blot Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for Flow Cyt or IP.
  • Target

    • Function

      Participates 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 specificity

      Expressed in testis, thymus, prostate and ovary and weakly expressed in lung, brain and kidney.
    • Sequence similarities

      Belongs 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 localization

      Cytoplasm. 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

    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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75814).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75814).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75814).

    • 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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab75814).

    References

    This product has been referenced in:

    • 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) »
    • 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) »
    See all 11 Publications for this product

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