- ALX-804-333-C100 - Enzo Life Sciences DIABLO antibody

Submitted references Degradation of Mcl-1 by granzyme B: implications for Bim-mediated mitochondrial apoptotic events.

Han J, Goldstein LA, Gastman BR, Froelich CJ, Yin XM, Rabinowich H
The Journal of biological chemistry 2004 May 21;279(21):22020-9

Tissue distribution of Diablo/Smac revealed by monoclonal antibodies.

Tikoo A, O'Reilly L, Day CL, Verhagen AM, Pakusch M, Vaux DL
Cell death and differentiation 2002 Jul;9(7):710-6

Multiple kinetics of mitochondrial cytochrome c release in drug-induced apoptosis.

Luetjens CM, Kögel D, Reimertz C, Düssmann H, Renz A, Schulze-Osthoff K, Nieminen AL, Poppe M, Prehn JH
Molecular pharmacology 2001 Nov;60(5):1008-19

Bcl-rambo, a novel Bcl-2 homologue that induces apoptosis via its unique C-terminal extension.

Kataoka T, Holler N, Micheau O, Martinon F, Tinel A, Hofmann K, Tschopp J
The Journal of biological chemistry 2001 Jun 1;276(22):19548-54

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Supportive validation

Submitted by: Enzo Life Sciences (provider)
Main image
Experimental details: Clone 10G7 can be used in Western blot to detect full-length Smac/DIABLO (~29kDa) as well as the mature form (~23kDa) resulting from N-terminal processing.Detection of endogenous and overexpressed DIABLO by Western blot by clone 10G7. The unprocessed form of DIABLO is detectable when overexpressed in 293T cells. When analysis of endogenous DIABLO (150 to 200µg total protein per lane) in healthy cells of human as well as mouse origin is performed, only the processed form can be detected, indicating that processing is not dependent on apoptosis. Recombinant processed mouse DIABLO serves as a positive control. Method: Recommended dilution: 1:500 to 1:1'000, incubation time with the antibody: 4-6 hours at room temperature, followed by incubation with an anti-rat HRP-conjugated antibody (Prod. No. ALX-211-052). Exposure time with (e.g. Pico chemoluminescence detection system Pierce): 10-30 min.

Supportive validation

Submitted by: Enzo Life Sciences (provider)
Main image
Experimental details: Clone 10G7 can be used in immunocytochemistry to follow the translocation of endogenous DIABLO. Smac/DIABLO (green) remains associated with the mitochondria in healthy human NT2 cells, as does cytochrome c (red). Upon UV irradiation both proteins are simultaneously released from the mitochondria and accumulate in the cytosol. . Method: 10G7 was used at a dilution of 1:100 but the optimal concentration needs to be determined individually. Cells were fixed with 4% PFA and permeabilized with 0.1-0.3% saponin, followed by incubation with an anti-rat-FITC conjugated antibody (Prod. No. ALX-211-051).

Supportive validation

Submitted by: Enzo Life Sciences (provider)
Main image
Experimental details: Clone 10G7 can be used in immunoprecipitation experiments, e.g. to study the interaction with IAPs. . Detection of endogenous and overexpressed DIABLO by immunoprecipitation with clone 10G7 followed by detection with a polyclonal rabbit antiserum to Smac.

ALX-804-333-C100

Antibody data