Name: Anti-Bcl-2 antibody [E17]
SKU: ab32124
Rating: 3 (14 reviews)

Anti-Bcl-2 antibody [E17] (ab32124) is a rabbit monoclonal antibody that is used to detect Bcl-2 in Western Blot, IP, IHC-P. Suitable for Human samples.

- Specificity confirmed with Bcl-2 knockout cell line validation

Related conjugates and formulations

ab185002
Carrier free
Anti-Bcl-2 antibody [E17] - BSA and Azide free
ab311800
Conjugated
Alexa Fluor® 594 Anti-Bcl-2 antibody [E17]
ab311028
Conjugated
Alexa Fluor® 488 Anti-Bcl-2 antibody [E17]
ab311154
Conjugated
Alexa Fluor® 647 Anti-Bcl-2 antibody [E17]
ab313083
Conjugated
Alexa Fluor® 568 Anti-Bcl-2 antibody [E17]
ab313277
Conjugated
Alexa Fluor® 555 Anti-Bcl-2 antibody [E17]
ab321578
Conjugated
Alexa Fluor® 750 Anti-Bcl-2 antibody [E17]

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (AB32124), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	Flow Cyt	WB	ICC/IF	IHC-P
Human	Tested	Not recommended	Tested	Not recommended	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes We recommend using ab182858 for murine samples.

Not recommended
Not recommended

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/250 - 1/500	Notes We do not recommend rat and mouse samples with IHC. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

15 products for Alternative Product

Target data

See full target information BCL2

Function

Suppresses apoptosis in a variety of cell systems including factor-dependent lymphohematopoietic and neural cells (PubMed:1508712, PubMed:8183370). Regulates cell death by controlling the mitochondrial membrane permeability (PubMed:11368354). Appears to function in a feedback loop system with caspases (PubMed:11368354). 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) (PubMed:11368354). Also acts as an inhibitor of autophagy: interacts with BECN1 and AMBRA1 during non-starvation conditions and inhibits their autophagy function (PubMed:18570871, PubMed:20889974, PubMed:21358617). May attenuate inflammation by impairing NLRP1-inflammasome activation, hence CASP1 activation and IL1B release (PubMed:17418785).

Alternative names

Apoptosis regulator Bcl-2, BCL2

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Clone number

E17

Purification technique

Affinity purification Protein A

Specificity

This antibody recognises Bcl-2. It does not cross-react with other Bcl-2 family members.

Bcl-2 has two isoforms, one is around 26kDa and the other is around 20kDa (PMID: 26009263, PMID: 10400666, PMID: 32377726).

Dissociation constant

3 x 10^-11 M

Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Abcam recommended secondaries - Goat Anti-Rabbit HRP (Goat Anti-Rabbit IgG H&L (HRP) ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077). Or search our wide range of secondary antibodies for use with your experiment.

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

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

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

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Bcl-2 also known as B-cell lymphoma 2 is an important protein involved in the regulation of apoptosis. It mechanically functions by inhibiting cell death and is a member of the Bcl-2 family of proteins. Bcl-2 has a molecular weight of approximately 26 kDa. It is expressed mainly in fetal and adult tissues including the immune system and is notably present in hematopoietic stem cells and various cell lines derived from lymphoid tissues. In laboratory practices scientists often assess Bcl-2 expression levels through techniques like western blotting using antibodies such as 10c4 for detection.

Biological function summary

The role of Bcl-2 extends to maintaining cell survival and balancing apoptotic signals. It functions as an anti-apoptotic molecule within the cell primarily involved in forming complexes with pro-apoptotic members of the Bcl-2 family like Bax and Bak. These interactions prevent mitochondrial membrane permeabilization which is an important step in apoptotic signal transduction. As an important regulator of apoptosis Bcl-2 helps in cellular homeostasis and appropriate immune responses under physiological conditions.

Pathways

Bcl-2 plays a critical role in the intrinsic or mitochondrial pathway of apoptosis. Within this pathway Bcl-2 interacts with other proteins such as cytochrome c and APAF-1 to inhibit apoptosis. This pathway involves a network of Bcl-2 family proteins where Bcl-2's anti-apoptotic role counteracts the pro-apoptotic activity of proteins like Bax. These interactions help fine-tune the apoptotic response to various internal stimuli maintaining cellular integrity and function.

Associated diseases and disorders

The dysregulation of Bcl-2 expression has been implicated in cancer and autoimmune diseases. Overexpression of Bcl-2 is associated with several types of cancer including lymphomas and breast cancer where its anti-apoptotic property leads to tumor cell survival and resistance to chemotherapy. Bcl-2 also connects to disorders such as autoimmune diseases due to its regulation of cell survival with proteins like Bim playing an opposite role in promoting apoptosis. Understanding Bcl-2's functionality within these contexts is essential for developing targeted therapies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

16 product images

Image from Némati F et al. PLoS One. 2014;9(1):e80836. Fig 2.; doi: 10.1371/journal.pone.0080836.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)
Bcl-2 expression determined by immunohistochemical analyses of the 4 human UM xenografts (between 3 to 5 tumors have been studied per condition).
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)
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.
Immunoprecipitation - Anti-Bcl-2 antibody [E17] (ab32124)
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.
All lanes: Immunoprecipitation - Anti-Bcl-2 antibody [E17] (ab32124)
Predicted band size: 26 kDa
Observed band size: 26 kDa
Western blot - Anti-Bcl-2 antibody [E17] (ab32124)
Lanes 1- 2: Merged signal (red and green). Green - ab32124 observed at 26 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
ab32124 was shown to react with BCL2 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human BCL2 knockout HeLa 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 (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (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.
All lanes: Western blot - Anti-Bcl-2 antibody [E17] (ab32124) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: BCL2 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human BCL2 knockout HeLa cell line (Human BCL2 knockout HeLa cell line ab255364)
Performed under reducing conditions.
Predicted band size: 26 kDa
Observed band size: 26 kDa
This image is courtesy of an anonymous customer review.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)
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.
Western blot - Anti-Bcl-2 antibody [E17] (ab32124)
Lanes 1 - 4: Merged signal (red and green). Green - ab32124 observed at 26 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - 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 Anti-GAPDH antibody [6C5] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging. 3% milk used as blocking agent.
All lanes: Western blot - Anti-Bcl-2 antibody [E17] (ab32124)
Lane 1: Wild-type HAP1 whole cell lysate at 20 µg
Lane 2: BCL2 knockout HAP1 whole cell lysate at 20 µg
Lane 3: HeLa whole cell lysate at 20 µg
Lane 4: THP-1 whole cell lysate at 20 µg
Predicted band size: 26 kDa
Western blot - Anti-Bcl-2 antibody [E17] (ab32124)
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-Bcl-2 antibody [E17] (ab32124) at 1/1000 dilution
Lane 1: MCF7 (human breast adenocarcinoma cell line) cell lysate at 20 µg
Lane 2: A431 (human epidermoid carcinoma cell line) cell lysate at 20 µg
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDa
Western blot - Anti-Bcl-2 antibody [E17] (ab32124)
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
MCF-7 cells express Bcl-2, while SK-BR-3 cells do not express Bcl-2 (PMID: 18430249)
All lanes: Western blot - Anti-Bcl-2 antibody [E17] (ab32124) at 1/200 dilution
Lane 1: MCF7 (human breast adenocarcinoma cell line) cell lysate at 10 µg
Lane 2: SK-BR-3 (human mammary gland adenocarcinoma cell line) cell lysate at 10 µg
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/50000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDa
Western blot - Anti-Bcl-2 antibody [E17] (ab32124)
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
All lanes: Western blot - Anti-Bcl-2 antibody [E17] (ab32124) at 1/10000 dilution
Lane 1: Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate at 20 µg
Lane 2: HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate at 20 µg
Lane 3: SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate at 20 µg
Secondary
All lanes: Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)
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. Goat Anti-Rabbit IgG H&L (HRP) 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.
Western blot - Anti-Bcl-2 antibody [E17] (ab32124)
All lanes: Western blot - Anti-Bcl-2 antibody [E17] (ab32124) at 1/1000 dilution
All lanes: Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
Predicted band size: 26 kDa
Observed band size: 26 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labelling Bcl-2 with unpurified ab32124 at 1/200 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human B cell lymphoma tissue labelling Bcl-2 with unpurified ab32124. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.
OI-RD Scanning - Anti-Bcl-2 antibody [E17] (ab32124)
We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody.
Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)
Tissue Microarrays for Anti-Bcl2 antibody [E17] using ab32124 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pretreated with heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins. The section was incubated with ab32124 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124)
Immunohistochemical analysis of formalin fixed paraffin embedded human lymphoma labelling Bcl-2 with ab32124 at a concentration of 1µg/ml.

The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.

ab32124 anti- Bcl-2 antibody [E17] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.

Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).