Description
Description: This assay employs a two-site sandwich ELISA to quantitate CYB5A in samples. An antibody specific for CYB5A has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any CYB5A present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for CYB5A is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of CYB5A bound in the initial step. The color development is stopped and the intensity of the color is measured.
Overview: Cytochrome b5 is a membrane bound hemoprotein which function as an electron carrier for several membrane bound oxygenases. It has two isoforms produced by alternative splicing. Isoform 1 is bound to the cytoplasmic side of the endoplasmic reticulum. It has a C-terminal transmembrane alpha-helix. Isoform 2 was found in cytoplasm. Defects in CYB5A are the cause of type IV hereditary methemoglobinemia.The derived amino acid sequence showed that the previous designation of several amino acids was in error. Liver and erythrocyte cytochrome b5 isoforms have the same amino acid sequence except at position 98, which is thr and pro for liver and erythrocyte proteins, respectively