Mao F C, Bremel R D
Endocrinology-Reproductive Physiology Program, University of Wisconsin, Madison 53706.
J Dairy Sci. 1991 Sep;74(9):2946-51. doi: 10.3168/jds.S0022-0302(91)78479-8.
Enzyme-linked immunosorbent assays for bovine alpha-lactalbumin and beta-lactoglobulin have been developed for measurements of serum and tissue culture samples. Either alpha-lactalbumin or beta-lactoglobulin antiserum was coated on ELISA plates. Biotinylated proteins were used in competition with unknown amount of proteins in samples. After unbound proteins were washed off, ExtrAvidin-peroxidase and tetramethylbenzidine were then used as a detection system. Crossreactivity of caseins or bovine serum albumin was less than .0001% in either alpha-lactalbumin or beta-lactoglobulin ELISA. Parallel curves from serial dilutions were obtained in serum and media samples. The additivity of alpha-lactalbumin and beta-lactoglobulin ELISA was validated in either serum or medium samples. The intraassay and interassay coefficients of variation for alpha-lactalbumin and beta-lactoglobulin ELISA were below 10% over 51 and 47 assays. The ELISA are useful in mammary gland biology studies for measuring milk whey protein in serum or culture media.
已开发出用于检测牛α-乳白蛋白和β-乳球蛋白的酶联免疫吸附测定法，用于测量血清和组织培养样品。将α-乳白蛋白或β-乳球蛋白抗血清包被在酶联免疫吸附测定板上。使用生物素化蛋白与样品中未知量的蛋白进行竞争。洗去未结合的蛋白后，然后使用抗生物素蛋白-过氧化物酶和四甲基联苯胺作为检测系统。在α-乳白蛋白或β-乳球蛋白酶联免疫吸附测定中，酪蛋白或牛血清白蛋白的交叉反应率均低于0.0001%。在血清和培养基样品中获得了系列稀释的平行曲线。α-乳白蛋白和β-乳球蛋白酶联免疫吸附测定的加和性在血清或培养基样品中得到验证。在51次和47次测定中，α-乳白蛋白和β-乳球蛋白酶联免疫吸附测定的批内和批间变异系数均低于10%。该酶联免疫吸附测定法在乳腺生物学研究中可用于测量血清或培养基中的乳清蛋白。
