Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Bak antibody [7D10] - BSA and Azide free (ab252251)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rat monoclonal [7D10] to Bak - BSA and Azide free
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Bak antibody [7D10] - BSA and Azide free
    See all Bak primary antibodies

  • Description

    Rat monoclonal [7D10] to Bak - BSA and Azide free

  • Host species

    Rat

  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: IP

  • Species reactivity

    Reacts with: Human

  • Immunogen

    The details of the immunogen for this antibody are not available.

  • Positive control

    • WB: HeLa, Jurkat, HEK-293, MCF7, T47D, Human heart, HAP1 lysates. IHC-P: Human breast carcinoma, colon carcinoma tissues.

  • General notes

    ab252251 is the carrier-free version of ab243151. 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

    ab252251 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.

    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.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.

  • Storage buffer

    Constituent: 100% PBS

  • Carrier free

    Yes
  • Concentration information loading...

  • Purity

    Ion Exchange Chromatography

  • Clonality

    Monoclonal

  • Clone number

    7D10

  • Isotype

    IgG2a

Images

  • Western blot - Anti-Bak antibody [7D10] - BSA and Azide free (ab252251)
    Western blot - Anti-Bak antibody [7D10] - BSA and Azide free (ab252251)
    All lanes : Anti-Bak antibody [7D10] (ab243151) at 1/5000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 2 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
    Lane 3 : HEK-293 (human embryonic kidney epithelial cell), whole cell lysate
    Lane 4 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate
    Lane 5 : T47D (human ductal breast epithelial tumor epithelial cell), whole cell lysate
    Lane 6 : Human heart tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution

    Predicted band size: 23 kDa
    Observed band size: 25,20 kDa why is the actual band size different from the predicted?



    This data was developed using ab243151, the same antibody clone in a different buffer formulation.

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

    The expression profile/ molecular weight  observed is consistent with what has been described in the literature (PMID: 12881569).

    This antibody reacts with an unidentifiable protein around 150kDa.

    Exposure time: 1 second.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bak antibody [7D10] - BSA and Azide free (ab252251)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bak antibody [7D10] - BSA and Azide free (ab252251)

    This data was developed using ab243151 the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue labeling Bak with ab243151 at 1/50 dilution followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Cytoplasmic staining on human breast carcinoma. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Western blot - Anti-Bak antibody [7D10] - BSA and Azide free (ab252251)
    Western blot - Anti-Bak antibody [7D10] - BSA and Azide free (ab252251)
    All lanes : Anti-Bak antibody [7D10] (ab243151) at 1/1000 dilution

    Lane 1 : Wild-type HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 2 : BAK1 knockout HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate
    Lane 3 : Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell), whole cell lysate
    Lane 4 : BAK1 knockout HAP1 (human chronic myelogenous leukemia near-haploid cell), whole cell lysate
    Lane 5 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rat IgG H&L (IRDye® 800CW) (ab253031) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) at 1/10000 dilution

    Predicted band size: 23 kDa
    Observed band size: 25 kDa why is the actual band size different from the predicted?



    This data was developed using ab243151, the same antibody clone in a different buffer formulation.

    Blocking and diluting buffer and concentration: Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS.

    Lanes 1-5: Merged signal (red and green). Green - ab243151 observed at 25kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.

    Lanes 1-2: ab243151 Anti-BAK1 antibody [7D10] was shown to react with BAK1 in HeLa cells in Western blot. Loss of signal was observed when BAK1 knockout sample was used. Wild-type and BAK1 knockout samples were subjected to SDS-PAGE.

    Lanes 3-4: ab243151 Anti-BAK1 antibody [7D10] was shown to react with BAK1 in HAP1 cells in Western blot. Loss of signal was observed when BAK1 knockout sample was used. Wild-type and BAK1 knockout samples were subjected to SDS-PAGE.

    ab243151 and Anti-GAPDH antibody [6C5] (ab8245) were incubated at 4? overnight at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat Anti-Rat IgG H&L (IRDye® 800CW) (ab253031) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bak antibody [7D10] - BSA and Azide free (ab252251)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bak antibody [7D10] - BSA and Azide free (ab252251)

    This data was developed using ab243151 the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue labeling Bak with ab243151 at 1/50 dilution followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Cytoplasmic staining on human colon carcinoma. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Anti-Bak antibody [7D10] - BSA and Azide free (ab252251)
    Anti-Bak antibody [7D10] - BSA and Azide free (ab252251)

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