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

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Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR17509] to Bcl-2
Suitable for: Flow Cyt (Intra), WB, IHC-P
Knockout validated
Reacts with: Mouse, Human

Carrier Free HRP

Product name

Anti-Bcl-2 antibody [EPR17509]
See all Bcl-2 primary antibodies
Description

Rabbit monoclonal [EPR17509] to Bcl-2
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), WB, IHC-Pmore details
Unsuitable for: ICC/IF
Species reactivity

Reacts with: Mouse, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human tonsil and thymus lysates; Jurkat, U-937, THP-1, HeLa, C2C12, WEHI -3 and NIH/3T3 whole cell lysates; Mouse brain, heart, kidney and spleen lysates; Human fetal kidney and fetal spleen lysates; Wild-type Hap1 cell lysate. IHC-P: Human tonsil tissue, Human endometrial cancer tissue, Mouse spleen tissue. Flow Cyt (intra): Jurkat cells.
General notes
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17509
Isotype

IgG
Research areas

Alternative Versions
- HRP Anti-Bcl-2 antibody [EPR17509] (ab209039)
- Anti-Bcl-2 antibody [EPR17509] - BSA and Azide free (ab219608)
Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human Bcl-2 protein (ab85156)
Related Products
- Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab182858 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
Flow Cyt (Intra)		1/250.
WB	(5)	1/2000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa).
IHC-P	(7)	1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Notes
Flow Cyt (Intra) 1/250.
WB 1/2000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Application notes

Is unsuitable for ICC/IF.

Function

Suppresses 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). May attenuate inflammation by impairing NLRP1-inflammasome activation, hence CASP1 activation and IL1B release (PubMed:17418785).
Tissue specificity

Expressed in a variety of tissues.
Involvement in disease

A 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 similarities

Belongs to the Bcl-2 family.
Domain

BH1 and BH2 domains are required for the interaction with BAX and for anti-apoptotic activity.
The BH4 motif is required for anti-apoptotic activity and for interaction with RAF1 and EGLN3.
The loop between motifs BH4 and BH3 is required for the interaction with NLRP1.
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. Ubiquitinated by SCF(FBXO10), leading to its degradation by the proteasome.
Cellular localization

Mitochondrion outer membrane. Nucleus membrane. Endoplasmic reticulum membrane.
Target information above from: UniProt accession P10415 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 596 Human
- Entrez Gene: 12043 Mouse
- Omim: 151430 Human
- SwissProt: P10415 Human
- SwissProt: P10417 Mouse
- Unigene: 150749 Human
- Unigene: 257460 Mouse
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
- Bcl-2 antibody
- Bcl2 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
- Protein phosphatase 1, regulatory subunit 50 antibody
see all

Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)

All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/2000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : BCL2 knockout HeLa cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

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

Lanes 1- 2: Merged signal (red and green). Green - ab182858 observed at 26 kDa. Red - 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 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 (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 (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 [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) (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 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 (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)

All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1 µg/ml

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
Observed band size: 26 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab182858 observed at 26 kDa. Red - 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 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 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.
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)

All lanes : 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: 8 seconds

Blocking/Diluting buffer 5% NFDM/TBST
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) (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 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)

All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/20000 dilution

Lane 1 : Human tonsil lysate
Lane 2 : Human thymus lysate
Lane 3 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
Lane 4 : U-937 (Human histiocytic lymphoma cells) whole cell lysate
Lane 5 : THP-1 (Human monocytic leukemia cells) whole cell lysate
Lane 6 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
Lane 7 : C2C12 (Mouse myoblast cell line) whole cell lysate
Lane 8 : WEHI-3 (Mouse leukemia cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : 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: 1 minute

Blocking and diluting buffer was 5% NFDM /TBST.
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)

All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/2000 dilution

Lane 1 : Mouse brain lysate
Lane 2 : Mouse heart lysate
Lane 3 : Mouse kidney lysate
Lane 4 : Mouse spleen lysate
Lane 5 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : 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: 3 minutes

Blocking and diluting buffer was 5% NFDM /TBST.
Western blot - Anti-Bcl-2 antibody [EPR17509] (ab182858)

All lanes : Anti-Bcl-2 antibody [EPR17509] (ab182858) at 1/2000 dilution

Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal spleen lysate

Lysates/proteins at 10 µg per lane.

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: 3 minutes

Blocking and diluting buffer was 5% NFDM /TBST.
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) (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 ab97051 at 1/500.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-Bcl-2 antibody [EPR17509] (ab182858)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab182858? Please let us know so that we can cite the reference in this datasheet.

ab182858 has been referenced in 287 publications.

Liu Z et al. Secreted TRAIL gene-modified adipose-derived stem cells exhibited potent tumor-suppressive effect in hepatocellular carcinoma cells. Immun Inflamm Dis 9:144-156 (2021). PubMed: 33156578
Xiao Y et al. Hsa_circ_0002483 regulates miR-758-3p/MYC axis to promote acute myeloid leukemia progression. Hematol Oncol 39:243-253 (2021). PubMed: 33283885
Zhou B et al. Protective Effects of Nicotinamide Riboside on H2O2-induced Oxidative Damage in Lens Epithelial Cells. Curr Eye Res 46:961-970 (2021). PubMed: 33297791
Shen H et al. Curcumin analog B14 has high bioavailability and enhances the effect of anti-breast cancer cells in vitro and in vivo. Cancer Sci 112:815-827 (2021). PubMed: 33316116
Wei X et al. Ultrasound-targeted microbubble destruction-mediated Galectin-7-siRNA promotes the homing of bone marrow mesenchymal stem cells to alleviate acute myocardial infarction in rats. Int J Mol Med 47:677-687 (2021). PubMed: 33416139

View all Publications for this product

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1-5 of 5 Abreviews or Q&A

Application

Western blot

Sample

Mouse Cell lysate - whole cell (CT26 cells)

Gel Running Conditions

Reduced Denaturing (4-12% Tris-Bis gel)

Loading amount

40 µg

Specification

CT26 cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Dec 08 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (primary AML cells)

Gel Running Conditions

Reduced Denaturing (4-12% Tris-Bis gel)

Loading amount

40 µg

Specification

primary AML cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Dec 08 2021

Application

Western blot

Sample

Human Cell lysate - whole cell (Glioblastoma cells)

Gel Running Conditions

Reduced Denaturing

Loading amount

10 µg

Treatment

4mg/ml bevacizumab for 48hrs

Specification

Glioblastoma cells

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 09 2020

Application

Western blot

Sample

Mouse Cell lysate - whole cell (MEFs)

Gel Running Conditions

Reduced Denaturing

Loading amount

30 µg

Specification

MEFs

Blocking step

BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Dr. Yonatan Feuermann

Verified customer

Submitted Jun 04 2020

Application

Western blot

Sample

Pig Tissue lysate - whole (Brain)

Gel Running Conditions

Non-reduced Denaturing (12% gel)

Loading amount

30 µg

Specification

Brain

Blocking step

Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jul 19 2019

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (287)

Customer reviews and Q&As

Western blot abreview for Anti-Bcl-2 antibody [EPR17509]

Western blot abreview for Anti-Bcl-2 antibody [EPR17509]

Western blot abreview for Anti-Bcl-2 antibody [EPR17509]

Western blot abreview for Anti-Bcl-2 antibody [EPR17509]

Western blot abreview for Anti-Bcl-2 antibody [EPR17509]

Post-translational
modifications