Description
Principle of the assay: human Diablo 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, A56-D239) 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.
Background: 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