Retinol binding protein (RBP) is a 21 kDa highly conserved, single-chain glycoprotein that has a hydrophobic pocket which binds retinol (vitamin A) in a 1:1 stoichiometry, which serves to not only solubilize retinol but also protect it from oxidation. RBP has also been shown to be a useful marker for renal function as it is totally filtered by the glomeruli and reabsorbed by proximal tubules. This has made urinary RBP (uRBP) a tool to study renal function in heart or kidney transplant recipients, type 1 and 2 diabetics, and in people exposed to uranium and other toxic metals from mining operations. Measurement of uRBP levels has also been useful in detection and characterization of diseases including hypertension and certain cancers, among other conditions.
The RBP Immunoassay Kit is designed to measure RBP levels in urine. This kit uses a native human RBP molecule as a standard, the concentration of which has been established via amino acid analysis. Standards or diluted samples are pipetted into a microtiter plate coated with a goat antibody to rabbit IgG. A RBP-HRP conjugate is added and the binding reaction initiated by addition of a RBP-specific rabbit polyclonal antibody. After a 60-minute incubation, the plate is washed and TMB substrate solution added to each well. After 30 minutes the intensity of the generated signal is read at 450 nm in a microtiter plate reader.
Human urine samples have been validated with this assay. Because RBP is a highly conserved protein we expect that RBP from other species will be read by this assay. The end user should evaluate the recovery of RBP from other species and sample types.
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