Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate LDHB in samples. An antibody specific for LDHB has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyLDHB present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for LDHB 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 LDHB bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:LDHB and peptidase B are linked (Santachiara et al., 1970) and both loci are on chromosome 12 (Chen et al., 1973). Kitamura et al. (1971) reported the first case of a complete deficiency of lactate dehydrogenase subunit H(B) in serum, saliva and erythrocytes of a 64-year-old male with mild diabetes. Study made on family members revealed low LDH activity in their serum also linked with decreased relative activity of the H4(B4) fraction. Based on the comparison of the calculated ratio of H to M subunits in normal and affected family members, it was hypothesized that the proband is homozygous while the abnormal family members are heterozygous, assuming a single gene is involved. Red cell metabolism in the proband was studied by Miwa et al. (1971); neither reticulocytosis nor hemolytic anemia was present.
