Revision 7

#2933Store 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, IF-IC, FC-FP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

12, 15, 23

Source/Isotype:

Rabbit IgG

UniProt ID:

#O43521

Entrez-Gene Id:

10018

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:50 - 1:250
Immunoprecipitation 1:200
IHC Leica Bond 1:100 - 1:400
Immunohistochemistry (Paraffin) 1:100 - 1:400
Immunofluorescence (Immunocytochemistry) 1:100 - 1:200
Flow Cytometry (Fixed/Permeabilized) 1:100 - 1:400

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 #26184.

Specificity / Sensitivity

Bim (C34C5) Rabbit mAb detects endogenous levels of total Bim (EL, L and S isoforms) protein.

Species Reactivity:

Human, Mouse, Rat

Species predicted to react based on 100% sequence homology

Monkey, Bovine, Dog, Pig

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro25 of Bim.

Background

Bim/Bod is a pro-apoptotic protein belonging to the BH3-only group of Bcl-2 family members including Bad, Bid, Bik, Hrk, and Noxa that contain a BH3 domain but lack other conserved BH1 or BH2 domains (1,2). Bim induces apoptosis by binding to and antagonizing anti-apoptotic members of the Bcl-2 family. Interactions have been observed with Bcl-2, Bcl-xL, Mcl-1, Bcl-w, Bfl-1, and BHRF-1 (1,2). Bim functions in regulating apoptosis associated with thymocyte negative selection and following growth factor withdrawal, during which Bim expression is elevated (3-6). Three major isoforms of Bim are generated by alternative splicing: BimEL, BimL, and BimS (1). The shortest form, BimS, is the most cytotoxic and is generally only transiently expressed during apoptosis. The BimEL and BimL isoforms may be sequestered to the dynein motor complex through an interaction with the dynein light chain and released from this complex during apoptosis (7). Apoptotic activity of these longer isoforms may be regulated by phosphorylation (8,9). Environmental stress triggers Bim phosphorylation by JNK and results in its dissociation from the dynein complex and increased apoptotic activity.

  1. O'Connor, L. et al. (1998) EMBO J 17, 384-95.
  2. Hsu, S.Y. et al. (1998) Mol Endocrinol 12, 1432-40.
  3. Bouillet, P. et al. (2002) Nature 415, 922-6.
  4. Whitfield, J. et al. (2001) Neuron 29, 629-43.
  5. Dijkers, P.F. et al. (2000) Curr Biol 10, 1201-4.
  6. Ley, R. et al. (2003) J Biol Chem 278, 18811-6.
  7. Puthalakath, H. et al. (1999) Mol Cell 3, 287-96.
  8. Lei, K. and Davis, R.J. (2003) Proc Natl Acad Sci U S A 100, 2432-7.
  9. Putcha, G.V. et al. (2003) Neuron 38, 899-914.

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) IF-IC: Immunofluorescence (Immunocytochemistry) 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.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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 7
#2933

Bim (C34C5) Rabbit mAb

Western Blotting Image 1: Bim (C34C5) Rabbit mAb Expand Image
使用 Bim (C34C5) Rabbit mAb 对 Raji、A20 和 RL 细胞提取物进行蛋白质印迹分析。
Western Blotting Image 2: Bim (C34C5) Rabbit mAb Expand Image
使用Bim (C34C5) Rabbit mAb #2933 and α-Tubulin (11H10) Rabbit mAb #2125,对经 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-), 或 SignalSilence® Bim siRNA I #6461,或SignalSilence® Bim siRNA II (+), 转染的HeLa细胞提取物进行蛋白质印迹分析Bim (C34C5) Rabbit mAb 确认 Bim 表达的沉默,而 α-Tubulin (11H10) Rabbit mAb 则用作上样和Bim siRNA异性对照。
Western Blotting Image 1: Bim (C34C5) Rabbit mAb Expand Image
使用 Bim (C34C5) Rabbit mAb #2933 对 Raji 细胞的裂解物 (1.0 mg/mL) 进行 Simple Western™ 分析。虚拟泳道视图(左图)显示一抗稀释比例为 1:50 和 1:250 时的三个靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:50(蓝线)和 1:250(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: Bim (C34C5) Rabbit mAb Expand Image
使用 Bim (C34C5) Rabbit mAb(泳道 1)或 Rabbit (DA1E) mAb IgG XP® Isotype Control(泳道 2)对 RL 细胞裂解物进行免疫沉淀法分析。使用 Bim (C34C5) Rabbit mAb 进行蛋白质印迹法分析。使用 Protein A (HRP Conjugate) #12291 进行二次检测。
Immunohistochemistry Image 1: Bim (C34C5) Rabbit mAb Expand Image
在 Leica® BOND Rx 上使用 Bim (C34C5) Rabbit mAb 对石蜡包埋的人结肠腺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: Bim (C34C5) Rabbit mAb Expand Image
使用 Bim (C34C5) Rabbit mAb 在 Leica® BOND Rx 上对石蜡包埋的人非小细胞肺癌组织进行免疫组织化学分析。
Immunohistochemistry Image 1: Bim (C34C5) Rabbit mAb Expand Image
使用 Bim (C34C5) Rabbit mAb 对石蜡包埋的人肺癌进行免疫组织化学分析。
Immunohistochemistry Image 2: Bim (C34C5) Rabbit mAb Expand Image
使用 Bim (C34C5) Rabbit mAb 对石蜡包埋的人淋巴瘤进行免疫组织化学分析。
Immunohistochemistry Image 3: Bim (C34C5) Rabbit mAb Expand Image
在对照肽(左)或抗原特异性肽(右)存在的情况下,使用 Bim (C34C5) Rabbit mAb 对石蜡包埋的人乳腺癌进行免疫组织化学分析。
Immunohistochemistry Image 4: Bim (C34C5) Rabbit mAb Expand Image
使用 Bim (C34C5) Rabbit mAb 对石蜡包埋的 4T1 同源基因肿瘤进行免疫组织化学分析。
Immunofluorescence Image 1: Bim (C34C5) Rabbit mAb Expand Image
使用 Bim Antibody(绿色)对 MCF-7 细胞进行共聚焦免疫荧光分析,显示与用 MitoTracker® Red CMXRos(红色)标记的线粒体共定位。蓝色伪彩 = DRAQ5® #4084(DNA 荧光染料)。
Flow Cytometry Image 1: Bim (C34C5) Rabbit mAb Expand Image
Flow cytometric analysis of fixed/permeabilized JeKo-1 cells (blue, negative) and Raji cells (green, positive) using Bim (C34C5) Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 用作二抗。
Flow Cytometry Image 2: Bim (C34C5) Rabbit mAb Expand Image
以浓度匹配的 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(虚线)作为对照,使用 Bim (C34C5) Rabbit mAb(实线)对 Raji 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 作为二抗。