Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E17] to Bcl-2 - BSA and Azide free
  • Suitable for: WB, IP, IHC-P
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Bcl-2 antibody [E17] - BSA and Azide free
    See all Bcl-2 primary antibodies

  • Description

    Rabbit monoclonal [E17] to Bcl-2 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HAP1, HeLa, THP-1 and MCF-7 cell lysates. IHC-P: Human DLBCL U2932, B-cell lymphoma, breast carcinoma and salivary gland tissue, and UM xenografts. IP: Jurkat cell lysate.

  • General notes

    Ab185002 is the carrier-free version of ab32124. 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

    ab185002 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.

Images

  • Western blot - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    Western blot - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    All lanes : Anti-Bcl-2 antibody [E17] (ab32124) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : BCL2 HeLa knockout 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 (ab32124).

      Lanes 1- 2: Merged signal (red and green). Green - ab32124 observed at 26 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

     ab32124 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. ab32124 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002) This image is courtesy of an anonymous Abreview.

    Formaldehyde-fixed, paraffin-embedded human DLBCL U2932 cell line xenograft tissue stained for Bcl-2 using ab32124 at 1/200 dilution in immunohistochemical analysis, followed by Goat anti-Rabbit IgG Alexa Fluor® 555.

    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 (ab32124).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002) Image from N?mati F et al. PLoS One. 2014;9(1):e80836. Fig 2.; doi: 10.1371/journal.pone.0080836.

    Bcl-2 expression determined by immunohistochemical analyses of the 4 human UM xenografts (between 3 to 5 tumors have been studied per condition).

    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 (ab32124).

  • Immunoprecipitation - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    Immunoprecipitation - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)

    ab32124 (purified) at 1/30 immunoprecipitating Bcl-2 in Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate (Lane 1). Lane 2 - PBS. For western blotting, a HRP-conjugated anti-rabbit IgG, specific to the non-reduced form of IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32124).

  • Western blot - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    Western blot - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    All lanes : Anti-Bcl-2 antibody [E17] (ab32124)

    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 per lane.

    Predicted band size: 26 kDa



    This data was developed using the same antibody clone in a different buffer formulation (ab32124).

    Lanes 1 - 4: Merged signal (red and green). Green - ab32124 observed at 26 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab32124 was shown to specifically react with BCL2 when BCL2 knockout samples were used. Wild-type and BCL2 knockout samples were subjected to SDS-PAGE.  Ab32124 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1000 dilution 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. 3% milk used as blocking agent.

  • Western blot - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    Western blot - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002) + MCF-7 (human breast carcinoma) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

    Predicted band size: 26 kDa


    Exposure time: 3 minutes


    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concentration: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human B-cell lymphoma tissue labelling Bcl-2 with purified ab32124 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32124).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Bcl-2 with unpurified ab32124 at 1/200 dilution.

    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 (ab32124).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human B cell lymphoma tissue labelling Bcl-2 with unpurified ab32124.

    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 (ab32124).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002) Image from Szyszko EA et al. Arthritis Res Ther. 2011 Jan 7;13(1):R2. Fig 5.; doi:10.1186/ar3220; 7 January 2011 Arthritis Research & Therapy 2011 13:R2.

    Immunohistochemical analysis of Human salivary glands taken from patients with primary Sjögren's syndrome, staining Bcl-2 with unpurified ab32124.


    Antigen retrieval was performed via heat mediation in a citrate buffer (pH 6). Sections were blocked using 2% BSA, 10% normal serum and permeabilized with 0.5% Triton X-100. Samples were incubated with primary antibody (1/100) for one hour at room temperature. An Alexa Fluor® 594-conjugated anti-rabbit IgG was used as the secondary antibody.
    N.B. Panels B and D are higher magnifications of panels A and C, respectively.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32124).

  • OI-RD Scanning - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    OI-RD Scanning - Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)

    Equilibrium disassociation constant (KD)

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32124).

  • Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)
    Anti-Bcl-2 antibody [E17] - BSA and Azide free (ab185002)

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