Diablo homolog, mitochondrial, also known as second mitochondria-derived activator of caspases(SMAC), is a protein that in humans is encoded by the DIABLO gene(direct IAP binding protein with low pI). It is mapped to 12q24.31. DIABLO can bind mammalian IAP homolog A(MIHA, or API3) and can also interact with MIHB, MIHC, and OpIAP, the baculoviral IAP. This gene encodes an inhibitor of apoptosis protein(IAP)-binding protein. The encoded mitochondrial protein enters the cytosol when cells undergo apoptosis, and allows activation of caspases by binding to inhibitor of apoptosis proteins. Over expression of the encoded protein sensitizes tumor cells to apoptosis. A mutation in this gene is associated with young-adult onset of nonsyndromic deafness-64.
Sample Type:Cell culture supernates, serum and plasma (heparin, EDTA)
Intended Use:For quantitative detection of human Diablo in cell culture supernates, serum and plasma (heparin, EDTA).
Sensitivity:<10pg/ml
Range:31.2-2000pg/ml
Specificity:Natural and recombinant human Diablo
Test Principle:This human Diablo BioAssay™ ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Diablo has been precoated onto 96-well plates. Standards(E.coli) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Diablo is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human Diablo amount of sample captured in plate.
Kit Components:96-well plate precoated with human Diablo antibody, 1Lyophilized recombinant human Diablo standard, 10ng/tube×2Biotinylated human Diablo antibody, 130ul (dilution 1:100)Avidin-Biotin-Peroxidase Complex (ABC), 130ul (dilution 1:100)Sample diluent buffer, 30mlAntibody diluent buffer, 12mlABC diluent buffer, 12mlTMB color developing agent, 10mlTMB stop solution, 10ml
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
