Revision 4

#15108

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, IP, IHC-P, IF-F, IF-IC, FC-FP

Reactivity:
H M R

Sensitivity:
Endogenous

MW (kDa):
21

Source/Isotype:
Rabbit IgG

UniProt ID:
#Q9NR28

Entrez-Gene Id:
56616

Product Usage Information

Application Dilution
Western Blotting 1:1000
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:500 - 1:2000
Immunofluorescence (Frozen) 1:100 - 1:400
Immunofluorescence (Immunocytochemistry) 1:100 - 1:400
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.

Specificity/Sensitivity

Smac/Diablo (D5S3R) Rabbit mAb recognizes endogenous levels of total Smac/Diablo protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human Smac/Diablo protein.

Background

Smac/Diablo is a 21 kDa mammalian mitochondrial protein that functions as a regulatory component during apoptosis (1,2). Upon mitochondrial stress, Smac/Diablo is released from mitochondria and competes with caspases for binding of IAPs (inhibitor of apoptosis proteins) (1,2). The interaction of Smac/Diablo with IAPs relieves the inhibitory effect of the IAPs on caspases (3,4). This interaction involves mainly the amino-terminal residues of Smac/Diablo with the BIR3 region of XIAP, supplemented with several other hydrophobic interactions between the helical structures of Smac/Diablo and other areas of BIR3 (5,6).

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 IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-F: Immunofluorescence (Frozen) 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.

Alexa Fluor is a registered trademark of Life Technologies Corporation.

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 4

Cell Signaling Technology Logo
Western blot analysis of extracts from various cell lines using Smac/Diablo (D5S3R) Rabbit mAb.
Western Blotting Image 1: Smac/Diablo (D5S3R) Rabbit mAb
Immunoprecipitation of Smac/Diablo from Jurkat cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Smac/Diablo (D5S3R) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot was performed using Smac/Diablo Mouse mAb #2954.
Immunoprecipitation Image 1: Smac/Diablo (D5S3R) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded colon carcinoma using Smac/Diablo (D5S3R) Rabbit mAb.
Immunohistochemistry Image 1: Smac/Diablo (D5S3R) 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 4

Cell Signaling Technology Logo
Immunohistochemical analysis of paraffin-embedded lung carcinoma using Smac/Diablo (D5S3R) Rabbit mAb.
Immunohistochemistry Image 2: Smac/Diablo (D5S3R) Rabbit mAb
Immunohistochemical analysis of paraffin-embedded prostate carcinoma using Smac/Diablo (D5S3R) Rabbit mAb.
Immunohistochemistry Image 3: Smac/Diablo (D5S3R) Rabbit mAb
Confocal immunofluorescent analysis within the white matter of mouse cerebellum using Smac/Diablo (D5S3R) Rabbit mAb #15108 (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 1: Smac/Diablo (D5S3R) 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 4

Cell Signaling Technology Logo
Confocal immunofluorescent tile scan of mouse brain using Smac/Diablo (D5S3R) Rabbit mAb #15108 (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue). Insets A – E are shown at higher resolution in subsequent figures.
Immunofluorescence Image 2: Smac/Diablo (D5S3R) Rabbit mAb
Inset A from mouse brain (cortex) using Smac/Diablo (D5S3R) Rabbit mAb #15108 (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 3: Smac/Diablo (D5S3R) Rabbit mAb
Inset B from mouse brain (striatum) using Smac/Diablo (D5S3R) Rabbit mAb #15108 (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 4: Smac/Diablo (D5S3R) 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 4

Cell Signaling Technology Logo
Inset C from mouse brain (hippocampus) using Smac/Diablo (D5S3R) Rabbit mAb #15108 (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 5: Smac/Diablo (D5S3R) Rabbit mAb
Inset D from mouse brain (thalamus) using Smac/Diablo (D5S3R) Rabbit mAb #15108 (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 6: Smac/Diablo (D5S3R) Rabbit mAb
Inset E from mouse brain (brainstem) using Smac/Diablo (D5S3R) Rabbit mAb #15108 (green) and GFAP (GA5) Mouse mAb #3670 (red). Samples were mounted in ProLong® Gold Antifade Reagent with DAPI #8961 (blue).
Immunofluorescence Image 7: Smac/Diablo (D5S3R) 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 4

Cell Signaling Technology Logo
Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Z-VAD (50 μM, 3 hr) and Staurosporine #9953 (1 μM, 3 hr; right), using Smac/Diablo (D5S3R) Rabbit mAb. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
Immunofluorescence Image 1: Smac/Diablo (D5S3R) Rabbit mAb
Flow cytometric analysis of Jurkat cells using Smac/Diablo (D5S3R) Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow Cytometry Image 1: Smac/Diablo (D5S3R) 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.