Smac/Diablo Rabbit Monoclonal Antibody for WB, IF/ICC, ELISA - Q9NR28

Core Product Specifications and Parameters

Biological Background: Smac/Diablo Function and Localization

• Diablo (Direct IAP binding protein with low pI), also known as Second mitochondria-derived activator of caspases (SMAC), is a pro-apoptotic protein that promotes apoptosis by activating caspases in the cytochrome c/Apaf-1/caspase-9 pathway.
• It functions by opposing the inhibitory activity of inhibitor of apoptosis proteins (IAP), specifically inhibiting BIRC6/BRUCE's binding to caspases. Related references: PMID:15200957 PMID:36758104 PMID:36758105
• Diablo attenuates XIAP/BIRC4 stability and apoptosis-inhibiting activity by promoting its ubiquitination and proteasomal degradation, and disrupts XIAP interaction with processed caspase-9 to activate caspase-3.
• The protein is localized to the mitochondrion and is released into the cytosol upon apoptotic stimuli.
• It is ubiquitously expressed, with highest levels in testis, and also highly expressed in heart, liver, kidney, spleen, prostate, and ovary; lower in brain, lung, thymus, and peripheral blood leukocytes.
• Alternative splicing produces multiple isoforms, and mutations in this gene are associated with non-syndromic deafness (DFNA64).
• Post-translational modifications include ubiquitin conjugation, and the protein contains a transit peptide for mitochondrial targeting.

Experimental Guidance and Technical Tips

• The immunogen corresponds to amino acids 56–239 of mature human Smac/Diablo, covering the functional region after mitochondrial processing; it is suitable for detecting the endogenous mature protein (~23 kDa processed form) and possibly the full-length precursor.
• For Western blotting (WB), consider using mitochondrial or whole-cell lysates; the predicted molecular weight of the full-length precursor is 27 kDa, while the mature form after transit peptide cleavage migrates at approximately 23 kDa. Optimize lysis and loading conditions accordingly.
• In immunofluorescence/immunocytochemistry (IF/ICC), mitochondrial localization may be observed under steady-state conditions; upon apoptosis induction, staining may shift to the cytosol.
• For ELISA, the antibody can be used to capture or detect recombinant or endogenous Smac/Diablo; pairing with a detection antibody against a non-overlapping epitope is recommended for optimal sensitivity.
• Cross-reactivity with mouse and rat is specified; when working with these species, validate the antibody in the relevant tissue or cell system to confirm reactivity and specificity.

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