Description
Principle of the assay: This assay employs a two-site sandwich ELISA to quantitative PLB2 in Human serum, plasma. An antibody specific for PLB2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any PLB2 present is bound by the immobilized antibody. After removing any unbound substances, a biotin - conjugated antibody specific for PLB2 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 PLB2 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Background: By Western blot analysis using an antibody against guinea pig phospholipase B (PLB), Maury et al. (2002) detected a 97-kD polypeptide in the soluble and membrane fractions of human epidermis that was enriched in the soluble fraction. Consistent with these results, they found that PLB activity was greatest in the soluble fraction. Trypsin had no effect on the molecular mass or enzymatic activity, suggesting that the protein they detected was already in an active form. Immunolocalization experiments revealed that PLB is expressed in the entire human epidermis with an accumulation in the dermoepidermis junction. In situ hybridization also indicated expression in the entire epidermis