Revision 7

#2933

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, 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 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 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.

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) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: Human M: Mouse R: Rat

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

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 7

Cell Signaling Technology Logo
Western blot analysis of extracts from Raji, A20 and RL cells using Bim (C34C5) Rabbit mAb.
Western Blotting Image 1: Bim (C34C5) Rabbit mAb
Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Bim siRNA I #6461 or SignalSilence® Bim siRNA II (+), using Bim (C34C5) Rabbit mAb #2933 and α-Tubulin (11H10) Rabbit mAb #2125. Bim (C34C5) Rabbit mAb confirms silencing of Bim expression and α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Bim siRNA.
Western Blotting Image 2: Bim (C34C5) Rabbit mAb
Simple Western™ analysis of lysates (1.0 mg/mL) from Raji cells using Bim (C34C5) Rabbit mAb #2933. The virtual lane view (left) shows three target bands (as indicated) at 1:50 and 1:250 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:50 (blue line) and 1:250 (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: Bim (C34C5) Rabbit 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 7

Cell Signaling Technology Logo
Immunoprecipitation of RL cell lysate using Bim (C34C5) Rabbit mAb (lane 1) or Rabbit (DA1E) mAb IgG XP® Isotype Control (lane 2). Western blot was performed using Bim (C34C5) Rabbit mAb. Protein A (HRP Conjugate) #12291 was used for secondary detection.
Immunoprecipitation Image 1: Bim (C34C5) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human colon adenocarcinoma using Bim (C34C5) Rabbit mAb performed on the Leica® BOND™ Rx.
Immunohistochemistry Image 1: Bim (C34C5) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human non-small cell lung carcinoma using Bim (C34C5) Rabbit mAb performed on the Leica® BOND™ Rx.
Immunohistochemistry Image 2: Bim (C34C5) Rabbit 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 7

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Bim (C34C5) Rabbit mAb.
Immunohistochemistry Image 1: Bim (C34C5) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human lymphoma using Bim (C34C5) Rabbit mAb.
Immunohistochemistry Image 2: Bim (C34C5) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Bim (C34C5) Rabbit mAb in the presence of control peptide (left) or antigen specific peptide (right).
Immunohistochemistry Image 3: Bim (C34C5) Rabbit 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 7

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded 4T1 syngeneic tumor using Bim (C34C5) Rabbit mAb.
Immunohistochemistry Image 4: Bim (C34C5) Rabbit mAb
Confocal immunofluorescent analysis of MCF-7 cells using Bim Antibody (green) showing colocalization with mitochondria that have been labeled with MitoTracker® Red CMXRos (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunofluorescence Image 1: Bim (C34C5) Rabbit mAb
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 was used as a secondary antibody.
Flow Cytometry Image 1: Bim (C34C5) Rabbit 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 7

Cell Signaling Technology Logo
Flow cytometric analysis of Raji cells using Bim (C34C5) Rabbit mAb (solid line) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed line). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow Cytometry Image 2: Bim (C34C5) Rabbit 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.