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    products/primary-antibodies/gsk3-beta-phospho-s9-antibody-epr2286y-ab75814.pdf

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RecombinantRabMAb

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

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  • SDS
Reviews (10)Q&A (1)References (81)

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Western blot - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
  • Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
  • Dot Blot - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
  • Western blot - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
  • Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2286Y] to GSK3 beta (phospho S9)
  • Suitable for: Dot blot, WB, IHC-P, ICC/IF
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

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

    Rabbit monoclonal [EPR2286Y] to GSK3 beta (phospho S9)
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

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

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR2286Y
  • Isotype

    IgG
  • Research areas

    • Neuroscience
    • Neurology process
    • Notch Pathway
    • Signal Transduction
    • Protein Phosphorylation
    • Ser / Thr Kinases
    • Other Kinases
    • Stem Cells
    • Signaling Pathways
    • Wnt
    • Cytoplasmic
    • Stem Cells
    • Signaling Pathways
    • Notch
    • Cytoplasmic
    • Cardiovascular
    • Atherosclerosis
    • Diabetes associated
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Other
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Integration of energy metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Integration of energy
    • Metabolism
    • Types of disease
    • Diabetes
    • Metabolism
    • Types of disease
    • Cancer
    • Metabolism
    • Types of disease
    • Heart disease
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

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

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • HeLa Ser/Thr Phosphatase-Inhibited Whole Cell Lysate Set: Calyculin-Treated and Vehicle-Treated (ab135395)
  • Related Products

    • Calyculin A, protein phosphatase inhibitor (ab141784)

Applications

The Abpromise guarantee

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
Dot blot
Use at an assay dependent concentration.
WB (7)
1/10000 - 1/20000. Detects a band of approximately 47 kDa (predicted molecular weight: 47 kDa).
IHC-P (1)
1/100 - 1/250. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF (2)
1/100 - 1/250.
Notes
Dot blot
Use at an assay dependent concentration.
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 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.
  • Target information above from: UniProt accession P49841 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 2932 Human
    • Omim: 605004 Human
    • SwissProt: P49841 Human
    • Unigene: 445733 Human
    • 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

    • Western blot - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
      Western blot - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
      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
      All lanes : 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
      Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)

      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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (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.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Dot Blot - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
      Dot Blot - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (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.

      Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

    • Western blot - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
      Western blot - Anti-GSK3 beta (phospho S9) antibody [EPR2286Y] (ab75814)
      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
      All lanes : goat anti-rabbit HRP at 1/2000 dilution

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

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

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols
    • Immunocytochemistry & immunofluorescence protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (81)

    Publishing research using ab75814? Please let us know so that we can cite the reference in this datasheet.

    ab75814 has been referenced in 81 publications.

    • Jiang H  et al. lncRNAS56464.1 as a ceRNA promotes the proliferation of fibroblast-like synoviocytes in experimental arthritis via the Wnt signaling pathway and sponges miR-152-3p. Int J Mol Med 47:N/A (2021). PubMed: 33448322
    • Luo P  et al. S-Allylmercaptocysteine improves alcoholic liver disease partly through a direct modulation of insulin receptor signaling. Acta Pharm Sin B 11:668-679 (2021). PubMed: 33777674
    • Yi Y  et al. Downregulation of DEC1 inhibits proliferation, migration and invasion, and induces apoptosis in ovarian cancer cells via regulation of Wnt/ß-catenin signaling pathway. Exp Ther Med 21:372 (2021). PubMed: 33732345
    • Wang S  et al. Relationship between long non-coding RNA PCAT-1 expression and gefitinib resistance in non-small-cell lung cancer cells. Respir Res 22:146 (2021). PubMed: 33980216
    • Ouyang Y  et al. Helicobacter pylori induces epithelial-mesenchymal transition in gastric carcinogenesis via the AKT/GSK3ß signaling pathway. Oncol Lett 21:165 (2021). PubMed: 33552283
    View all Publications for this product

    Customer reviews and Q&As

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

    1-10 of 11 Abreviews or Q&A

    Western blot abreview for Anti-GSK3 beta (phospho S9) antibody [EPR2286Y]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Melanoma)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    40 µg
    Specification
    Melanoma
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 06 2023

    Western blot abreview for Anti-GSK3 beta (phospho S9) antibody [EPR2286Y]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (CT26 cells)
    Gel Running Conditions
    Reduced Denaturing (4-20% Tris Glycin gel)
    Loading amount
    40 µg
    Specification
    CT26 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 01 2022

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-GSK3 beta (phospho S9) antibody [EPR2286Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Human Hippocampus Section)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate pH6
    Permeabilization
    No
    Specification
    Human Hippocampus Section
    Blocking step
    Bloxall as blocking agent for 20 minute(s) · Concentration: 100% · Temperature: 20°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Dec 06 2021

    Western blot abreview for Anti-GSK3 beta (phospho S9) antibody [EPR2286Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (SHSY5Y cells)
    Gel Running Conditions
    Reduced Denaturing (4-12% Tris-Bis gel)
    Loading amount
    40 µg
    Specification
    SHSY5Y cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Aug 07 2021

    Western blot abreview for Anti-GSK3 beta (phospho S9) antibody [EPR2286Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (LNCaP - prostate cancer cells)
    Gel Running Conditions
    Reduced Denaturing (4-15% SDS PAGE)
    Loading amount
    30 µg
    Specification
    LNCaP - prostate cancer cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Armen Petrosyan

    Verified customer

    Submitted Dec 29 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-GSK3 beta (phospho S9) antibody [EPR2286Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Armen Petrosyan

    Verified customer

    Submitted Dec 19 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-GSK3 beta (phospho S9) antibody [EPR2286Y]

    Excellent
    Abreviews
    Abreviews
    abreview image
    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%
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Armen Petrosyan

    Verified customer

    Submitted Dec 19 2017

    Western blot abreview for Anti-GSK3 beta (phospho S9) antibody [EPR2286Y]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Liver)
    Gel Running Conditions
    Reduced Denaturing (12% separating gel, 5% stacking gel)
    Loading amount
    80 µg
    Treatment
    50 mg/kg body weight Chloroquine diphosphate
    Specification
    Liver
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Dec 23 2016

    Western blot abreview for Anti-GSK3 beta (phospho S9) antibody [EPR2286Y]

    Poor
    Abreviews
    Abreviews
    abreview image
    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
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    DR. Karine Thibault

    Verified customer

    Submitted Nov 10 2014

    Question

    which antibody is best for detecting phospho protein?

    Read More

    Abcam community

    Verified customer

    Asked on Aug 15 2012

    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.

    Use our products? Submit an Abreview. Earn rewards!
    https://www.abcam.com/abreviews

    Read More

    Abcam Scientific Support

    Answered on Aug 15 2012

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