Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20445] to BTK - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP
  • Reacts with: Human

Overview

  • Product name

    Anti-BTK antibody [EPR20445] - BSA and Azide free
    See all BTK primary antibodies

  • Description

    Rabbit monoclonal [EPR20445] to BTK - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human

  • Immunogen

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

  • Positive control

    • IHC-P: Human tonsil tissue.

  • General notes

    Ab227812 is the carrier-free version of ab208937. 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

    ab227812 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)

    Immunohistochemical analysis of paraffin-embedded human colon tissue labeling BTK with ab208937 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Strong cytoplasmic staining on lymphoid nodule in human colon (PMID: 25433814). 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 (ab208937).

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

  • Immunocytochemistry/ Immunofluorescence - Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)
    Immunocytochemistry/ Immunofluorescence - Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)

    Immunofluorescent analysis of 100% methanol-fixed Ramos (human Burkitt's lymphoma cell line) and Jurkat (human T cell leukemia cell line from peripheral blood) cells labeling BTK with ab208937 at 1/100 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 Ramos cell line.

    Negative control: Jurkat cell line (PMID: 24759210).

    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 (ab208937).

  • Flow Cytometry - Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)
    Flow Cytometry - Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)

    Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized Jurkat (human T cell leukemia cell line from peripheral blood) (left panel) and Ramos (human Burkitt's lymphoma cell line) (right panel) cell lines labeling BTK with ab208937 at 1/600 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.

    Negative control: Jurkat cell line (PMID: 24759210).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab208937).

  • Immunoprecipitation - Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)
    Immunoprecipitation - Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)

    BTK was immunoprecipitated from 0.35 mg of Ramos (human Burkitt's lymphoma cell line) whole cell lysate with ab208937 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab208937 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Ramos whole cell lysate 10 µg (Input). 

    Lane 2: ab208937 IP in Ramos whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab208937 in Ramos 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 (ab208937).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling BTK with ab208937 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Strong cytoplasmic staining in mantle zone and weaker cytoplasmic staining in the germinal center of human tonsil (PMID: 25433814). 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 (ab208937).

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

  • Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)
    Anti-BTK antibody [EPR20445] - BSA and Azide free (ab227812)

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