Revision 8

#15071Store at -20C

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:

WB, 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 Information

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.

Species Reactivity:

Human

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).

  1. Murphy, K.M. et al. (2000) Cell Death Differ 7, 102-11.
  2. Zhu, L. et al. (1999) J Biol Chem 274, 33267-73.
  3. Maundrell, K. et al. (1997) J Biol Chem 272, 25238-42.
  4. Yamamoto, K. et al. (1999) Mol Cell Biol 19, 8469-78.
  5. Ling, Y.H. et al. (1998) J Biol Chem 273, 18984-91.
  6. Huang, S.T. and Cidlowski, J.A. (2002) FASEB J 16, 825-32.
  7. Deng, X. et al. (2001) J Biol Chem 276, 23681-8.

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

WB: 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 M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

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

Revision 8
#15071

Bcl-2 (124) Mouse mAb

Western Blotting Image 1: Bcl-2 (124) Mouse mAb Expand Image
使用 Bcl-2 (124) Mouse mAb #15071(上图)或 β-actin (13E5) Rabbit mAb #4970(下图)对对照型 HeLa 细胞 (lane 1) 或 Bcl-2 敲除型 HeLa 细胞 (lane 2) 的提取物进行蛋白印迹分析。Bcl-2 敲除型 HeLa 细胞中没有信号,这证实了该抗体对 Bcl-2 的特异性。
Western Blotting Image 2: Bcl-2 (124) Mouse mAb Expand Image
使用 Bcl-2 (124) Mouse mAb(上)和 β-Actin (D6A8) Rabbit mAb #8457(下),对不同细胞系提取物进行蛋白质印迹分析。KARPAS 细胞系来源:University of Cambridge 的 Abraham Karpas 博士。
Western Blotting Image 1: Bcl-2 (124) Mouse mAb Expand Image
使用 Bcl-2 (124) Mouse mAb #15071 对 Jurkat 细胞的裂解物 (1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。

Immunoprecipitation Image 1: Bcl-2 (124) Mouse mAb Expand Image
使用 Mouse (G3A1) mAb IgG1 Isotype Control #5415(泳道 2)或 Bcl-2 (124) Mouse mAb(泳道 3),对 RL 细胞提取物 Bcl-2 进行免疫沉淀分析。泳道 1 是 10% 输入对照。使用 Bcl-2 (D55G8) Rabbit mAb (Human Specific) #4223 进行蛋白质印迹分析。
Immunohistochemistry Image 1: Bcl-2 (124) Mouse mAb Expand Image
在 Leica® BOND Rx 上使用 Bcl-2 (124) mouse mAb 对石蜡包埋的人乳腺导管癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: Bcl-2 (124) Mouse mAb Expand Image
在 Leica® BOND Rx 上使用 Bcl-2 (124) mouse mAb 对石蜡包埋的人非霍奇金氏淋巴瘤进行免疫组织化学分析。
Immunohistochemistry Image 3: Bcl-2 (124) Mouse mAb Expand Image
在 Leica® BOND Rx 上使用 Bcl-2 (124) mouse mAb 对石蜡包埋的人卵巢浆液性癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 1: Bcl-2 (124) Mouse mAb Expand Image
使用 Bcl-2 (124) Mouse mAb 对石蜡包埋的人乳腺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: Bcl-2 (124) Mouse mAb Expand Image
使用 Bcl-2 (124) Mouse mAb 对石蜡包埋的人肺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 3: Bcl-2 (124) Mouse mAb Expand Image
使用 Bcl-2 (124) Mouse mAb 对石蜡包埋的 RL(阳性,左图)和 HT-29(阴性,右图)细胞沉淀物进行免疫组织化学分析。
Immunohistochemistry Image 4: Bcl-2 (124) Mouse mAb Expand Image
使用 Bcl-2 (124) Mouse mAb 对石蜡包埋的人扁桃体细胞进行免疫组织化学分析。
Flow Cytometry Image 1: Bcl-2 (124) Mouse mAb Expand Image
使用 Bcl-2 (124) Mouse mAb(实线)或浓度匹配的 Mouse (G3A1) mAb IgG1 Isotype Control #5415(虚线)对 HT-29 细胞(蓝色)和 RL 细胞(绿色)进行流式细胞分析。Anti-mouse IgG (H+L),F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4408 作为二抗。