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Technical Data Sheet

SM3591

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Smac/DIABLO (C-terminal region)
Mouse Monoclonal IgG1
Price
Size
Species Reactivity
MW

$245
100 μl
Hu, Rt, Ms
23 kDa

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Western blot analysis of Smac/DIABLO expression in A431 cell lysate. The blots were probed with mouse monoclonal Smac/DIABLO (C-terminal region) at 1:500 (lane 1) or 1:2000 (lane 2).

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Enlarge

Immunocytochemical labeling of Smac/Diablo in aldehyde-fixed and NP-40-permeabilized A431 cells. The cells were labeled with mouse monoclonal Smac/Diablo (C-terminal region) antibody, then the antibody was detected using appropriate secondary antibody conjugated to DyLight® 594. The cells were counterstained with Sytox green to label nuclei.

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Application
Dilution


ELISA
1:2000


ICC
1:100


WB
1:1000



End user should determine optimal dilution for their particular applications and experiments.Western blot membranes were incubated with diluted antibody in 5% non-fat milk, PBS, 0.04% Tween20 for 1hour at room temperature.
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Background
X linked inhibitor of apoptosis (XIAP) is a member of the inhibitor of apoptosis (IAP) family of proteins, and is involved in regulating programmed cell death by caspase inhibition. SMAC/DIABLO (second mitochondria derived activator of caspase/direct IAP binding protein with low PI) is expressed in a variety of tissues, and has been identified as a negative regulator of XIAP. A functionally active nine-residue peptide derived from the N terminus of Smac/DIABLO interacts with the baculovirus IAP repeat 3 domain of XIAP. Smac/DIABLO promotes caspase activation in the cytochrome c/Apaf-1/caspase-9 pathway by binding to IAPs and removing their inhibitory activity. Smac/DIABLO is normally a mitochondrial protein but is released into the cytosol when cells undergo apoptosis.


Background References
Du, C. et al. (2000) Cell 102:33.
Liu, Z. et al. (2000) Nature 408:1004.
Verhagen, A.M. et al. (2000) Cell 102:43.
Immunogen
Clone M359 was generated from a recombinant protein that included amino acid residues within the C-terminal region of human Smac/DIABLO. This sequence has high homology with similar regions in rat and mouse Smac/DIABLO.
Buffer and Storage
Mouse monoclonal antibody purified with protein A chromatography is supplied in 100µl phosphate-buffered saline, 50% glycerol, 1 mg/ml BSA, and 0.05% sodium azide. Store at –20°C. Stable for 1 year.
Specificity
The antibody detects a 23 kDa* band corresponding to the molecular weight of Smac/DIABLO on SDS-PAGE immunoblots of A431 cells.

*All molecular weights (MW) are confirmed by comparison to Bio-Rad Rainbow Markers and to western blot mobilities of known proteins with similar MW.
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