Overview

  • Product nameAnti-Bcl-2 antibody [E17]
    See all Bcl-2 primary antibodies
  • Description
    Rabbit monoclonal [E17] to Bcl-2
  • SpecificityThe antibody recognises Bcl-2. It does not cross-react with other Bcl-2 family members.
  • Tested applicationsWB, IHC-P, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Bcl-2 aa 50-150.
    Database link: P10415

  • Positive control
    • WB: MCF-7, A431, Jurkat, HeLa, SH-SY5Y and NIH/3T3 cell lysates. IHC-P: human B-cell lymphona, lung adenocarcinoma and breast carcinoma tissues. ICC/IF: MCF-7 cells. Flow Cyt: Jurkat cells. IP: Jurkat cell lysate.
  • General notes

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    A trial size is available for this product.

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Properties

Applications

Our Abpromise guarantee covers the use of ab32124 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/1000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa).
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

We do not recommend rat and mouse samples with IHC.

ICC/IF 1/100.
Flow Cyt 1/200.
IP 1/50.

Target

  • FunctionSuppresses apoptosis in a variety of cell systems including factor-dependent lymphohematopoietic and neural cells. Regulates cell death by controlling the mitochondrial membrane permeability. Appears to function in a feedback loop system with caspases. Inhibits caspase activity either by preventing the release of cytochrome c from the mitochondria and/or by binding to the apoptosis-activating factor (APAF-1).
  • Tissue specificityExpressed in a variety of tissues.
  • Involvement in diseaseA chromosomal aberration involving BCL2 has been found in chronic lymphatic leukemia. Translocation t(14;18)(q32;q21) with immunoglobulin gene regions. BCL2 mutations found in non-Hodgkin lymphomas carrying the chromosomal translocation could be attributed to the Ig somatic hypermutation mechanism resulting in nucleotide transitions.
  • Sequence similaritiesBelongs to the Bcl-2 family.
  • DomainThe BH4 motif is required for anti-apoptotic activity and for interaction with RAF1 and EGLN3.
  • Post-translational
    modifications
    Phosphorylation/dephosphorylation on Ser-70 regulates anti-apoptotic activity. Growth factor-stimulated phosphorylation on Ser-70 by PKC is required for the anti-apoptosis activity and occurs during the G2/M phase of the cell cycle. In the absence of growth factors, BCL2 appears to be phosphorylated by other protein kinases such as ERKs and stress-activated kinases. Phosphorylated by MAPK8/JNK1 at Thr-69, Ser-70 and Ser-87, wich stimulates starvation-induced autophagy. Dephosphorylated by protein phosphatase 2A (PP2A).
    Proteolytically cleaved by caspases during apoptosis. The cleaved protein, lacking the BH4 motif, has pro-apoptotic activity, causes the release of cytochrome c into the cytosol promoting further caspase activity.
    Monoubiquitinated by PARK2, leading to increase its stability.
  • Cellular localizationMitochondrion outer membrane. Nucleus membrane. Endoplasmic reticulum membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • Apoptosis regulator Bcl 2 antibody
    • Apoptosis regulator Bcl-2 antibody
    • Apoptosis regulator Bcl2 antibody
    • AW986256 antibody
    • B cell CLL/lymphoma 2 antibody
    • B cell leukemia/lymphoma 2 antibody
    • B cell lymphoma 2 antibody
    • Bcl 2 antibody
    • Bcl-2 antibody
    • Bcl2 antibody
    • BCL2 protein antibody
    • BCL2_HUMAN antibody
    • C430015F12Rik antibody
    • D630044D05Rik antibody
    • D830018M01Rik antibody
    • Leukemia/lymphoma, B-cell, 2 antibody
    • Oncogene B-cell leukemia 2 antibody
    • PPP1R50 antibody
    • Protein phosphatase 1, regulatory subunit 50 antibody
    see all

Anti-Bcl-2 antibody [E17] images

  • All lanes : Anti-Bcl-2 antibody [E17] (ab32124) at 1/1000 dilution (puriifed)

    Lane 1 : MCF-7 cell lysate
    Lane 2 : A431 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • All lanes : Anti-Bcl-2 antibody [E17] (ab32124) at 1/10000 dilution (purified)

    Lane 1 : Jurkat cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : SH-SY5Y cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Anti-Bcl-2 antibody [E17] (ab32124) at 1/1000 dilution (purified) + NIH/3T3 cell lysate at 20 µg

    Secondary
    Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Anti-Bcl-2 antibody [E17] (ab32124) at 1/1000 dilution (unpurified) + Jurkat cell lysate

    Predicted band size : 26 kDa
    Observed band size : 26 kDa
  • 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.

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung adenocarcinoma tissue labelling Bcl-2 with unpurified ab32124.

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

  • 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.

  • Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling Bcl-2 with unpurified ab32124 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/500) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were used.

    Control: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

  • Immunocytochemistry/Immunofluorescence analysis of MCF-7 cells labelling Bcl-2 with purified ab32124 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/500) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Overlay histogram showing Jurkat cells stained with unpurified ab32124 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32124, 1/100 dilution) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in Jurkat cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • ab32124 (purified) at 1/30 immunoprecipitating Bcl-2 in Jurkat 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.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References for Anti-Bcl-2 antibody [E17] (ab32124)

This product has been referenced in:
  • Némati F  et al. Targeting Bcl-2/Bcl-XL induces antitumor activity in uveal melanoma patient-derived xenografts. PLoS One 9:e80836 (2014). IHC-P ; Mouse . Read more (PubMed: 24454684) »
  • Xiao Q  et al. BEX1 promotes imatinib-induced apoptosis by binding to and antagonizing BCL-2. PLoS One 9:e91782 (2014). IP ; Human . Read more (PubMed: 24626299) »

See all 14 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Sample Human Cell (MCF-7)
Specification MCF-7
Permeabilization Yes - Triton X-100
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Sep 16 2014

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - 70% methanol
Sample Human Cell (MCF-7)
Specification MCF-7
Gating Strategy live cells
Preparation Cell harvesting/tissue preparation method: Cell dissociation buffer
Sample buffer: Enzyme free
Username

Abcam user community

Verified customer

Submitted Sep 02 2014

Application Western blot
Loading amount 50000 cells
Gel Running Conditions Reduced Denaturing (10% gel)
Sample Human Cell lysate - whole cell (MCF-7, MDA-MB-231)
Specification MCF-7, MDA-MB-231
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Aug 22 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - other (Jurkat T-cells)
Total protein in input 20 µg
Specification Jurkat T-cells
Immuno-precipitation step Protein A/G
Username

Dr. Jonathan Rud

Verified customer

Submitted Feb 19 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Jurkat T-cells)
Loading amount 25 µg
Specification Jurkat T-cells
Gel Running Conditions Reduced Denaturing (8% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Jonathan Rud

Verified customer

Submitted Feb 11 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (U937)
Specification U937
Fixation Leucoperm Reagent A
Permeabilization Yes - Leucoperm Reagent B
Gating Strategy Live Cells
Username

Abcam user community

Verified customer

Submitted Sep 09 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (U937 and PBMC)
Loading amount 4e+006 cells
Specification U937 and PBMC
Gel Running Conditions Reduced Denaturing (10% Gel)
Blocking step 2% Milk with 1% BSA as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C
Username

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Verified customer

Submitted Sep 03 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (jurkat)
Loading amount 15 µg
Specification jurkat
Gel Running Conditions Reduced Denaturing (12)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Karen Rees-Unwin

Verified customer

Submitted Jul 29 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (B16 Melanoma)
Loading amount 25 µg
Specification B16 Melanoma
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Mr. John Kelly

Verified customer

Submitted Dec 28 2007

The antibody recognizes both the alpha and beta forms of Bcl-2.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"