Name: Anti-Bcl-2 antibody [EPR17509]
SKU: ab182858
Rating: 5 (13 reviews)

Anti-Bcl-2 antibody [EPR17509] ab182858 is a rabbit monoclonal antibody that is used in Bcl-2 western blotting, IHC and flow cytometry. Suitable for human and mouse samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with BCL2 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR17509 is cited in over 880 publications

Images

Western blot - Anti-Bcl-2 antibody [EPR17509] (AB182858), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2
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

	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Tested	Not recommended	Tested	Tested
Mouse	Tested	Not recommended	Expected	Tested

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/2000	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 Mouse, Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/250	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Human	Dilution info 1/500	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

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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
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: EPR17509
Purification technique: Affinity purification Protein A
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

Anti-Bcl-2 antibody [EPR17509] (ab182858) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in Flow Cyt (Intra), IHC-P and WB.

Anti-Bcl-2 antibody [EPR17509] (ab182858) was first used in a scientific publication in 2016 and has been cited over 632 times in peer reviewed journals. It's performance in Western Blot in human and mouse samples is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-Bcl-2 antibody [EPR17509] (ab182858) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

The specificity of Anti-Bcl-2 antibody [EPR17509] (ab182858) has been confirmed by Western Blot testing in Bcl-2 knockout HeLa cells (ab263752).

Anti-Bcl-2 antibody [EPR17509] (ab182858) has 13 independent reviews from customers.

Anti-Bcl-2 antibody [EPR17509] (ab182858) specifically detects Bcl-2 (UniProt ID: P10415; Molecular weight: 27kDa) and is sold in 100 µL and 1 mL selling sizes.

Conjugation-ready, carrier free format available for antibody clone EPR17509 - Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free ab219608.

Antibody clone EPR17509 is also available pre-conjugated to a variety of labels for your convenience - HRP (HRP Anti-Bcl-2 antibody [EPR17509] ab209039).

B-cell lymphoma 2 (Bcl-2) is a key protein in oncology, known for its role in regulating apoptosis, or programmed cell death. It promotes cell survival by inhibiting pro-apoptotic proteins, thereby preventing apoptosis and contributing to the development and progression of various cancers.

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

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.

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:
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12 product images

Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)
Lanes 1- 2: Merged signal (red and green). Green - ab182858 observed at 26 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (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 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. ab182858 and Anti-GAPDH antibody [6C5] - Loading Control (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 (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 [EPR17509] (ab182858) at 1/2000 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
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)
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) (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 Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cytometry (Intracellular) - Anti-Bcl-2 antibody [EPR17509] (ab182858)
Intracellular 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 (Rabbit IgG, monoclonal [EPR25A] - 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.
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)
Lanes 1 - 4: Merged signal (red and green). Green - ab182858 observed at 26 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - 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 Anti-GAPDH antibody [6C5] - Loading Control 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 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.
All lanes: Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1 µg/mL
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
Observed band size: 26 kDa
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)
Blocking/Diluting buffer 5% NFDM/TBST
All lanes: Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/10000 dilution
Lane 1: NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 20 µg
Lane 2: WEHI-3 (mouse leukemia cell line) whole cell lysate at 20 µg
Lane 3: Mouse hippocampus at 10 µg
Lane 4: Mouse heart at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG H&L (HRP) at 1/2000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDa
Exposure time: 8s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)
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) (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 Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)
Blocking and diluting buffer was 5% NFDM /TBST.
All lanes: Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/20000 dilution
Lane 1: Human tonsil lysate at 20 µg
Lane 2: Human thymus lysate at 20 µg
Lane 3: Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg
Lane 4: U-937 (Human histiocytic lymphoma cells) whole cell lysate at 20 µg
Lane 5: THP-1 (Human monocytic leukemia cells) whole cell lysate at 20 µg
Lane 6: HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 7: C2C12 (Mouse myoblast cell line) whole cell lysate at 20 µg
Lane 8: WEHI-3 (Mouse leukemia cell line) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 26 kDa
Observed band size: 26 kDa
Exposure time: 1min
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)
Blocking and diluting buffer was 5% NFDM /TBST.
All lanes: Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/2000 dilution
Lane 1: Mouse brain lysate at 10 µg
Lane 2: Mouse heart lysate at 10 µg
Lane 3: Mouse kidney lysate at 10 µg
Lane 4: Mouse spleen lysate at 10 µg
Lane 5: NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate at 10 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 26 kDa
Observed band size: 26 kDa
Exposure time: 3min
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)
Blocking and diluting buffer was 5% NFDM /TBST.
All lanes: Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/2000 dilution
Lane 1: Human fetal kidney lysate at 10 µg
Lane 2: Human fetal spleen lysate at 10 µg
Secondary
All lanes: Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 26 kDa
Observed band size: 26 kDa
Exposure time: 3min
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)
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) (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 Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)
Immunohistochemical analysis of formalin fixed paraffin embedded human spleen labelling Bcl-2 with ab182858 at a concentration of 0.86µg/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.
ab182858 anti-Bcl-2 antibody [EPR17509] 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)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [EPR17509] (ab182858)
Immunohistochemical analysis of formalin fixed paraffin embedded human spleen labelling Bcl-2 with ab182858 at a concentration of 0.32µg/ml. The immunostaining was performed on a Leica Biosystems BOND ® RX instrument with a Bond™ Polymer Refine Detection kit. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution 2) for 20mins.
ab182858 anti-Bcl-2 antibody [EPR17509] was incubated for 15mins at room temperature. Sections were counterstained with Hematoxylin. 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)