Overview

  • Product name

    Anti-GSK3 beta (phospho S9) antibody
    See all GSK3 beta primary antibodies
  • Description

    Rabbit polyclonal to GSK3 beta (phospho S9)
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IHC-P, WBmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat, Sheep, Rabbit, Cow, Dog, Pig, Chimpanzee, Zebrafish, Macaque monkey, Gorilla
  • Immunogen

    Synthetic peptide corresponding to Human GSK3 beta aa 1-100 conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab129226)

  • Positive control

    • This antibody gave a positive signal in the following whole cell lysates: HeLa; Jurkat; A549. It also gave a positive result in MCF7 cell line. This antibody gave a positive result in IHC in the following FFPE tissue: Human breast adenocarcinoma

Properties

Applications

Our Abpromise guarantee covers the use of ab107166 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 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).

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

  • All lanes : Anti-GSK3 beta (phospho S9) antibody (ab107166) at 1 µg/ml (Milk blocked - 5%)

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Immunizing peptide at 1 µg/ml
    Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate with Immunizing peptide at 1 µg/ml
    Lane 6 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate with Immunizing peptide at 1 µg/ml

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 47 kDa
    Observed band size: 47 kDa
    Additional bands at: 125 kDa, 28 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 12 minutes
  • IHC image of GSK3 beta (phospho S9) staining in Human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab107166, 1µg/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.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

  • ICC/IF image of ab107166 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 (ab107166, 1µg/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. This antibody also gave a positive result in 4% formaldehyde fixed (10 min) MCF7 cells at 1µg/ml.

References

This product has been referenced in:

  • Marathe S  et al. Notch signaling in response to excitotoxicity induces neurodegeneration via erroneous cell cycle reentry. Cell Death Differ 22:1775-84 (2015). Read more (PubMed: 25822340) »
See 1 Publication for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Application
Western blot
Sample
Rat Cell lysate - whole cell (Rat hepatocytes)
Gel Running Conditions
Reduced Denaturing (8% SDS-PAGE)
Loading amount
40 µg
Specification
Rat hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Nov 19 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Rat hepatocytes)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
Rat hepatocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde

Dr. Armen Petrosyan

Verified customer

Submitted May 24 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (LNCaP - prostate cancer cells)
Permeabilization
Yes - 0.2% Triton X-100 in PBS
Specification
LNCaP - prostate cancer cells
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Fixative
Formaldehyde

Dr. Armen Petrosyan

Verified customer

Submitted Apr 10 2018

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

Dr. Karine Thibault

Verified customer

Submitted Nov 10 2014

Question
Answer

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 hesitate to contact us if you need any more advice or information.

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Question
Answer

Code de remise : ****
Date d’expiration : JJ MM AAAA

Désolé pour la non-disponibilité des autres anti-GSK3 beta (phospho Ser9).

Je suis ravi que vous souhaitiez participerà notre offre et testerab107166 chez la souris (100% homologie entre l'immunogène et la protéine cible). Le code de réduction correspondant est ******. Ce code de réduction est utilisable sous 120 jours et pour l’achat d’un autre anticorps primaire.

Vous devez dans un premier temps soumettre une Abreview avec vos résultatschez la sourisen notant ce code dans la section « Additional notes ». Ce code sera ainsi activé et vous pourrez par la suite contacter notre service clientèle par téléphone pour passer votre prochaine commande, en vous munissant des informations suivantes : code de réduction, numéro de commande de ab107166.

Nous restons à votre disposition pour plus d’informations concernant cette offre promotionnelle. Dans l’attente de recevoir votre Abreview nous vous souhaitons bonne chance dans vos recherches.

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