Revision 8

#15071

Store at -20C

CST Logo
Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Applications:
W, W-S, IP, IHC-Bond, IHC-P, FC-FP

Reactivity:
H

Sensitivity:
Endogenous

MW (kDa):
26

Source/Isotype:
Mouse IgG1

UniProt ID:
#P10415

Entrez-Gene Id:
596

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50
IHC Leica Bond 1:800 - 1:3200
Immunohistochemistry (Paraffin) 1:400 - 1:1600
Flow Cytometry (Fixed/Permeabilized) 1:200 - 1:800

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

For a carrier-free (BSA and azide free) version of this product see product #17447

Specificity/Sensitivity

Bcl-2 (124) Mouse mAb recognizes endogenous levels of total Bcl-2 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly47 of human Bcl-2 protein.

Background

Bcl-2 exerts a survival function in response to a wide range of apoptotic stimuli through inhibition of mitochondrial cytochrome c release (1). It has been implicated in modulating mitochondrial calcium homeostasis and proton flux (2). Several phosphorylation sites have been identified within Bcl-2, including Thr56, Ser70, Thr74, and Ser87 (3). It has been suggested that these phosphorylation sites may be targets of the ASK1/MKK7/JNK1 pathway and that phosphorylation of Bcl-2 may be a marker for mitotic events (4,5). Mutation of Bcl-2 at Thr56 or Ser87 inhibits its anti-apoptotic activity during glucocorticoid-induced apoptosis of T lymphocytes (6). Interleukin-3 and JNK-induced Bcl-2 phosphorylation at Ser70 may be required for its enhanced anti-apoptotic functions (7).

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

W: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-Bond: IHC Leica Bond IHC-P: Immunohistochemistry (Paraffin) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: Human

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

SignalStain is a registered trademark of Cell Signaling Technology, Inc.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品 , (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 8

Cell Signaling Technology Logo
Western blot analysis of extracts from control HeLa cells (lane 1) or Bcl-2 knockout HeLa cells (lane 2) using Bcl-2 (124) Mouse mAb #15071 (upper), or β-actin (13E5) Rabbit mAb #4970 (lower). The absence of signal in the Bcl-2-knockout HeLa cells confirms specificity of the antibody for Bcl-2.
Western Blotting Image 1: Bcl-2 (124) Mouse mAb
Western blot analysis of extracts from various cell lines using Bcl-2 (124) Mouse mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). KARPAS cell Line source: Dr Abraham Karpas at the University of Cambridge.
Western Blotting Image 2: Bcl-2 (124) Mouse mAb
Simple Western™ analysis of lysates (1 mg/mL) from Jurkat cells using Bcl-2 (124) Mouse mAb #15071. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ ​​​​​​​ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.

Western Blotting Image 1: Bcl-2 (124) Mouse mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 8

Cell Signaling Technology Logo
Immunoprecipitation of Bcl-2 from RL cell extracts using Mouse (G3A1) mAb IgG1 Isotype Control #5415 (lane 2) or Bcl-2 (124) Mouse mAb (lane 3). Lane 1 is 10% input. Western blot was performed using Bcl-2 (D55G8) Rabbit mAb (Human Specific) #4223.
Immunoprecipitation Image 1: Bcl-2 (124) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human ductal breast carcinoma using Bcl-2 (124) mouse mAb performed on the Leica® BOND™ Rx.
Immunohistochemistry Image 1: Bcl-2 (124) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human non-Hodgkin’s lymphoma using Bcl-2 (124) mouse mAb performed on the Leica® BOND™ Rx. 
Immunohistochemistry Image 2: Bcl-2 (124) Mouse mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 8

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human ovarian clear cell carcinoma using Bcl-2 (124) mouse mAb performed on the Leica® BOND™ Rx.
Immunohistochemistry Image 3: Bcl-2 (124) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Bcl-2 (124) Mouse mAb.
Immunohistochemistry Image 1: Bcl-2 (124) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Bcl-2 (124) Mouse mAb.
Immunohistochemistry Image 2: Bcl-2 (124) Mouse mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.

Revision 8

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded RL (positive, left) and HT-29 (negative, right) cell pellets using Bcl-2 (124) Mouse mAb.
Immunohistochemistry Image 3: Bcl-2 (124) Mouse mAb
Immunohistochemical analysis of paraffin-embedded human tonsil using Bcl-2 (124) Mouse mAb.
Immunohistochemistry Image 4: Bcl-2 (124) Mouse mAb
Flow cytometric analysis of HT-29 cells (blue) and RL cells (green) using Bcl-2 (124) Mouse mAb (solid lines) or a concentration-matched Mouse (G3A1) mAb IgG1 Isotype Control #5415 (dashed lines). Anti-mouse IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 was used as a secondary antibody.
Flow Cytometry Image 1: Bcl-2 (124) Mouse mAb
Orders: 877-616-CELL (2355) [email protected] Support: 877-678-TECH (8324) [email protected] Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.