Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

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 - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] - BSA and Azide free

  • Description

    Rabbit monoclonal [EPR18814-102] to GSK3 beta + GSK3 alpha - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IPmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

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

  • Positive control

    • IHC-P: Mouse testis tissue.

  • General notes

    Ab226169 is the carrier-free version of ab185141. 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

    ab226169 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Images

  • Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] - BSA and Azide free (ab226169)
    Western blot - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] - BSA and Azide free (ab226169)
    All lanes : Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] (ab185141) at 1/5000 dilution

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

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 47, 52 kDa



    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185141).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] - BSA and Azide free (ab226169)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] - BSA and Azide free (ab226169)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185141).

    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] - BSA and Azide free (ab226169)
    Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] - BSA and Azide free (ab226169)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185141).

  • Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] - BSA and Azide free (ab226169)
    Immunocytochemistry/ Immunofluorescence - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] - BSA and Azide free (ab226169)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185141).

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

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185141).

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

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185141).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] - BSA and Azide free (ab226169)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GSK3 beta + GSK3 alpha antibody [EPR18814-102] - BSA and Azide free (ab226169)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab185141).

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

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

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