Enzyme-linked immunosorbent assays for bovine a-lactalbumin and p..lactoglobulin have been developed for measurements of serum and tissue culture samples.Either a-lactalbumin or ~ 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, Ex- trAvidin-peroxidase and tetramethylbenzidine were then used as a detection system.Crossreactivity of caseins or b0vine serum albumin was less than .0001% in either a-lactalbumin or p.. lactoglobulin ELISA.Parallel curves from serial dilutions were obtained in serum and media samples.The additivity of a-lactalbumin and p..lactoglobulin ELISA was validated in either serum or medium samples.The intraassay and interassay coefficients of variation for alactalbumin and j3-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.(Key words: enzyme-linked immunosorbent assay, a-lactalbumin, p..lactoglobulin)Abbreviation key: DAB = diaminobenzidine, EIA =enzyme immunoassay, RIA =radioimmunoassay, TMB = tetramethylbenzidine.~dex of mammary gland function in response eIther to hormonal regulation in bovine explant culture (1, 8) or as an index of udder development and function (12).
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Mao et al. (1991) studied this question.
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