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    gsk3-beta--gsk3-alpha-antibody-epr18814-102-ab185141.pdf

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Cell Biology Cell Cycle Kinases/Phosphatases Cdks
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
  • Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
  • Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
  • Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
  • Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
  • Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
  • Flow Cytometry (Intracellular) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
  • Immunoprecipitation - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
  • Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18814-102] to GSK3 beta + GSK3 alpha
  • Suitable for: IHC-P, IP, ICC/IF, WB, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102]
  • Description

    Rabbit monoclonal [EPR18814-102] to GSK3 beta + GSK3 alpha
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, IP, ICC/IF, WB, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: His-tagged mouse GSK3 beta (aa218-420) recombinant protein; His-tagged mouse GSK3 alpha (aa330-490) recombinant protein; Human fetal brain, fetal heart and fetal kidney lysates; NIH/3T3, HeLa, A549, RAW 264.7 and PC-12 whole cell lysates; Mouse and rat heart lysates. IHC-P: Mouse and rat testis tissues. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): NIH/3T3 cells. IP: NIH/3T3 whole cell lysate.
  • 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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 0.05% BSA, 40% Glycerol (glycerin, glycerine), PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR18814-102
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Cell Cycle
    • Kinases/Phosphatases
    • Cdks
    • 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
    • Apoptosis
    • 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
    • Heart disease

Associated products

  • Alternative Versions

    • Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] - BSA and Azide free (ab226169)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Related Products

    • Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab185141 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
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

This antibody is not recommended for IHC in human.

IP
1/30.
ICC/IF
1/150.
WB
1/5000. Detects a band of approximately 47, 52 kDa (predicted molecular weight: 47, 52 kDa).
Flow Cyt (Intra)
1/60.
Notes
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

This antibody is not recommended for IHC in human.

IP
1/30.
ICC/IF
1/150.
WB
1/5000. Detects a band of approximately 47, 52 kDa (predicted molecular weight: 47, 52 kDa).
Flow Cyt (Intra)
1/60.

Target

  • Cellular localization

    GSK3 beta: 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.
  • Database links

    • Entrez Gene: 2931 Human
    • Entrez Gene: 2932 Human
    • Entrez Gene: 56637 Mouse
    • Entrez Gene: 606496 Mouse
    • Entrez Gene: 50686 Rat
    • Entrez Gene: 84027 Rat
    • Omim: 605004 Human
    • Omim: 606784 Human
    • SwissProt: P49840 Human
    • SwissProt: P49841 Human
    • SwissProt: Q2NL51 Mouse
    • SwissProt: Q9WV60 Mouse
    • SwissProt: P18265 Rat
    • SwissProt: P18266 Rat
    • Unigene: 445733 Human
    • Unigene: 466828 Human
    • Unigene: 294664 Mouse
    • Unigene: 394930 Mouse
    • Unigene: 10426 Rat
    • Unigene: 36807 Rat
    see all

Images

  • Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: GSK3 alpha knockout HAP1 whole cell lysate (20 µg)
    Lane 3: GSK3 beta whole cell lysate (20 µg)
    Lane 4: HeLa whole cell lysate (20 µg)
    Lane 5: Hek293 whole cell lysate (20 µg)

    Lanes 1 - 5: Merged signal (red and green). Green - ab185141 observed at 47/52 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab185141 was shown to specifically react with GSK3 alpha and GSK3 beta in wild-type HAP1 cells as signal was lost in GSK3 alpha and GSK3 beta knockout cells. Wild-type and GSK3 alpha and GSK3 beta knockout samples were subjected to SDS-PAGE. ab185141 and ab18058 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

    Immunohistochemical analysis of paraffin-embedded mouse testis tissue labeling GSK3 beta + GSK3 alpha with ab185141 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on mouse testis (PMID: 22792253). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    All lanes : Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141) at 1/1000 dilution

    Lane 1 : His-tagged mouse GSK3 beta (aa218-420) recombinant protein
    Lane 2 : His-tagged mouse GSK3 alpha (aa330-490) recombinant protein

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Developed using the ECL technique.

    Predicted band size: 47, 52 kDa


    Exposure time: 1 second


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    All lanes : Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141) at 1/5000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

    Developed using the ECL technique.

    Predicted band size: 47, 52 kDa
    Observed band size: 47,52 kDa why is the actual band size different from the predicted?



     

    Exposure time : Lane 1: 30 seconds; Lanes 2 and 3: 3 minutes.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    All lanes : Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141) at 1/5000 dilution

    Lane 1 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate
    Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : A549 (human lung carcinoma cell line) whole cell lysate
    Lane 4 : RAW 264.7 (mouse macrophage cell line transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 5 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 6 : Mouse heart lysate
    Lane 7 : Rat heart lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 47, 52 kDa
    Observed band size: 47,52 kDa why is the actual band size different from the predicted?



     

    Exposure time : Lanes 1 and 2:10 seconds; Lane 3: 1 minute; Lanes 4 and 5: 5 seconds; Lane 6:3 seconds; Lane 7:10 seconds.

    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

    Immunohistochemical analysis of paraffin-embedded rat testis tissue labeling GSK3 beta + GSK3 alpha with ab185141 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on rat testis (PMID: 22792253). Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling GSK3 beta + GSK3 alpha with ab185141 at 1/150 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HeLa cells.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling GSK3 beta + GSK3 alpha with ab185141 at 1/150 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on NIH/3T3 cells.

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

  • Flow Cytometry (Intracellular) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    Flow Cytometry (Intracellular) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed,90% methanol-permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cell line labeling GSK3 beta + GSK3 alpha with ab185141 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody. 

     

     

  • Immunoprecipitation - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    Immunoprecipitation - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

    GSK3 beta + GSK3 alpha was immunoprecipitated from 0.35 mg of NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate with ab185141 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab185141 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: NIH/3T3 whole cell lysate 10 µg (Input). 

    Lane 2: ab185141 IP in NIH/3T3 whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab185141 in NIH/3T3 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 seconds.

  • Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)
    Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (1)

Publishing research using ab185141? Please let us know so that we can cite the reference in this datasheet.

ab185141 has been referenced in 1 publication.

  • Fan C  et al. Myocardial protection by nanomaterials formulated with CHIR99021 and FGF1. JCI Insight 5:N/A (2020). PubMed: 32453715

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