Antibodies, Proteins, Kits and Reagents for Life Science

348 441 ₸ Product size

100 µl 348 441 ₸

Order now and get it on Tuesday March 02, 2021

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [E17] to Bcl-2
Suitable for: WB, IHC-P, IP
Knockout validated
Reacts with: Human

Product name

Anti-Bcl-2 antibody [E17]
See all Bcl-2 primary antibodies
Description

Rabbit monoclonal [E17] to Bcl-2
Host species

Rabbit
Specificity

Anti-Bcl-2 antibody [E17] (ab32124) recognises Bcl-2. It does not cross-react with other Bcl-2 family members.

Tested Applications & Species

Application	Species
IHC-P	Human
IP	Human
WB	Human

See all applications and species data

Immunogen

Synthetic peptide within Human Bcl-2 aa 50-150. The exact sequence is proprietary.
Database link: P10415
Positive control
- WB: MCF7, A431, Jurkat, HeLa, THP-1 and SH-SY5Y cell lysates; Wild type HAP1 whole cell lysate. IHC-P: Human B-cell lymphona and breast carcinoma tissues; Human UM xenografts; Human salivary glands; Human DLBCL U2932 cell line xenograft tissue. IP: Jurkat whole cell lysate (ab7899).
General notes
Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor^® 488 (ab150077).

See other anti-rabbit secondary antibodies that can be used with this 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 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.
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.

Form

Liquid
Storage instructions

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

K_D = 3.00 x 10 ^-11 M
Learn more about K_D
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

E17
Isotype

IgG
Research areas

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. 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 : Anti-Bcl-2 antibody [E17] (ab32124) at 1/1000 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 - ab32124 observed at 26 kDa. Red - 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 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 (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 (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 [E17] (ab32124) This image is courtesy of an anonymous Abreview.

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.
See Abreview
Western blot - Anti-Bcl-2 antibody [E17] (ab32124)

All lanes : Anti-Bcl-2 antibody [E17] (ab32124)

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

Lanes 1 - 4: Merged signal (red and green). Green - ab32124 observed at 26 kDa. Red - 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 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 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. 3% milk used as blocking agent.
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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Bcl-2 antibody [E17] (ab32124) Image from Némati F et al., PLoS One. 2014;9(1):e80836. Fig 2.; doi: 10.1371/journal.pone.0080836. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

Bcl-2 expression determined by immunohistochemical analyses of the 4 human UM xenografts (between 3 to 5 tumors have been studied per condition).
Western blot - Anti-Bcl-2 antibody [E17] (ab32124)

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

Lane 1 : MCF7 (human breast adenocarcinoma cell line) cell lysate
Lane 2 : A431 (human epidermoid carcinoma cell line) cell lysate

Lysates/proteins at 20 µg per lane.

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

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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.
Western blot - Anti-Bcl-2 antibody [E17] (ab32124)

All lanes : Anti-Bcl-2 antibody [E17] (ab32124) at 1/200 dilution

Lane 1 : MCF7 (human breast adenocarcinoma cell line) cell lysate
Lane 2 : SK-BR-3 (human mammary gland adenocarcinoma cell line) cell lysate

Lysates/proteins at 10 µg per lane.

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

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)
Western blot - Anti-Bcl-2 antibody [E17] (ab32124)

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

Lane 1 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 3 : SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate

Lysates/proteins at 20 µg per lane.

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

Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Western blot - Anti-Bcl-2 antibody [E17] (ab32124)

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

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.
OI-RD Scanning - Anti-Bcl-2 antibody [E17] (ab32124)

Equilibrium disassociation constant (K_D)

Click here to learn more about K_D
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.
Anti-Bcl-2 antibody [E17] (ab32124)

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