Biliverdin Reductase, Recombinant, Rat (BVR)

Biliverdin reductase (BVR), Heme oxygenase (HO) and UDP-glucuronosyltransferases (UGTs) form the major erythrocytic heme catabolic pathway in humans and most mammalian species. BVR is an oxireductase which catalyzes the reduction of the gama-meso bridge of biliverdin to bilirubin. Recent studies indicate that BVR is a phosphoprotein and that phosphorylation is essential to the activity of BVR to reduce biliverdin to bilirubin. These studies also found that the enzyme is autophosphorylated and that phosphorylation is reversible. In mammals bilirubin can be both a neurotoxin and an antioxidant depending on its ratio to protein and concentration; has immunomodulatory properties; induces Aryl hydrocarbon receptor-mediated activation of cytochrome P450 1a1 expression and inhibits protein phosphorylation (1). BVR is unique among all enzymes characterized to date because BVR has dual pH/dual cofactor requirements. The reductase is highly conserved in its primary structure and molecular properties and uses NADH in the acidic range (peak activity at pH~6.7), whereas NADPH is utilized in the basic range (peak activity at pH ~8.7) (2). BVR is encoded by a single copy gene and the ~1.6kb BVR message is abundantly expressed in kidney, spleen, liver and brain as well as at lower levels in the thymus and minimal levels being detected in testis (3). The human BVR was found to resolve into four isoelectric zones and two molecular mass forms (40.7kDa and 39.6kDa) in the liver whereas variants were detected in the kidney (4). Protein: Rat kidney biliverdin reductase; The protein is produced using an E. coli expression system (3). Applications: Suitable for use in Western Blot analysis and should be resuspended with an appropriate amount of SDS-PAGE sample buffer for this purpose. For in house use, the product is resuspended to 1ug/ul with 20ul 1X sample buffer (e.g. Laemmli, U.K., (1970) Nature 227: 680-685) and is then diluted further to 0.01ug/ul with 1X sample buffer. In a 15 well mini-blot system, 50ng/well gives a good signal when using antibody at 1:200 as the primary antibody and a goat anti-rabbit IgG alkaline phosphatase conjugate at 1:1,000 for detection. Storage and Stability: May be stored at 4 degrees C for short-term only. For long-term storage, aliquot to avoid repeated freezing and thawing and freeze at -70 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Aliquots are stable for at least 12 months.