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

    Does not react with

    Rat
  • 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
    • Jurkat cell lysate and human breast carcinoma.
  • General notes

    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.

    This product is available conjugated to DyLight® 488 - see ab139926.

    A trial size is available for this product.

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 - 1/10000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa).
IHC-P Use at an assay dependent concentration.

We do not recommend rat and mouse samples with IHC.

ICC/IF 1/200.
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

  • Immunofluorescent staining of MCF-7 cells using anti-Bcl-2 RabMAB(green) diluted 1/100 and counter stain with anti-tubulin RabMAB(red) shows staining in the cytoplasm.

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

    Predicted band size : 26 kDa
    Observed band size : 26 kDa
  • Immunohistochemical analysis of paraffin-embedded human breast carcinoma using ab32124 at 1/200 dilution.Immunohistochemical analysis of paraffin-embedded human breast carcinoma using ab32124 at 1/200 dilution.
  • ab32124 showing positive staining in Lung adenocarcinoma tissue.

  • ab32124 showing positive staining in B cell lymphoma tissue.

  • Overlay histogram showing Jurkat cells stained with 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.

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

  • 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"