Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR17509] to Bcl-2 - BSA and Azide free
  • Suitable for: Flow Cyt, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free
    See all Bcl-2 primary antibodies

  • Description

    Rabbit monoclonal [EPR17509] to Bcl-2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, WB, IHC-Pmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab85156)

  • Positive control

    • WB: Human tonsil lysate, Human thymus lysate; Jurkat, U-937, THP-1, HeLa, HAP1, C2C12, WEHI -3 and NIH/3T3 whole cell lysates; Mouse brain lysate, Mouse heart lysate, Mouse kidney lysate, Mouse spleen lysate, Human fetal kidney lysate, Human fetal spleen lysate. IHC-P: Human tonsil tissue, Human endometrial cancer tissue, Mouse spleen tissue. ICC/IF: Jurkat cells. Flow Cyt: Jurkat cells.

  • General notes

    Ab219608 is the carrier-free version of ab182858. 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

    ab219608 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Images

  • Western blot - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)
    Western blot - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)
    All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/2000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : BCL2 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 26 kDa
    Observed band size: 26 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab182858).

      Lanes 1- 2: Merged signal (red and green). Green - ab182858 observed at 26 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab182858 was shown to react with Bcl-2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255364 (knockout cell lysate ab263752) was used. Wild-type HeLa and BCL2 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab182858 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)
    Flow Cytometry - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)

    Flow cytometric analysis of 4% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood) cells labeling Bcl-2 with ab182858 at 1/250 (red) compared with a rabbit monoclonal IgG isotype control (ab172730) (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody (blue)). Goat anti rabbit IgG (FITC) at 1/500 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 (ab182858).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling Bcl-2 with ab182858 at 1/1000 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.

    Cytoplasm, nuclear membrane and nucleus staining on lymphocytes of Mouse spleen tissue is observed.

    Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary antibody followed by ab97051 at 1/500.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182858).

  • Western blot - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)
    Western blot - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)
    All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1 µg/ml

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : BCL2 knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : THP-1 whole cell lysate

    Lysates/proteins at 20 µg/ml per lane.

    Predicted band size: 26 kDa
    Observed band size: 26 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab182858).

    Lanes 1 - 4: Merged signal (red and green). Green - ab182858 observed at 26 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab182858 was shown to specifically react with BCL2 when BCL2 knockout samples were used. Wild-type and BCL2 knockout samples were subjected to SDS-PAGE. Ab182858 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 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-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)

    Immunohistochemical analysis of paraffin-embedded Human endometrial cancer tissue labeling Bcl-2 with ab182858 at 1/1000 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.

    Cytoplasm, nuclear membrane and nucleus staining on lymphocytes and cancer cells of  Human endometrial cancer  tissue is observed.

    Counter stained with Hematoxylin.
    Negative control: Used PBS instead of primary antibody followed by ab97051 at 1/500.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182858).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)

    Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Bcl-2 with ab182858 at 1/1000 followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500.

    Cytoplasm, nuclear membrane and nucleus staining on lymphocytes of Human tonsil tissue is observed.

    Counter stained with Hematoxylin.

    Negative control: Used PBS instead of primary antibody followed by ab97051 at 1/500.

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab182858).

  • Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)
    Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)

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