Several methods are now available for the immunoassay of insulin in plasma. Of these, the double antibody technique of Hales & Randle (1963) is especially convenient. However, in studies of plasma insulin concentrations in the sheep with method C of Hales & Randle (1963), plasma samples assayed undiluted consistently yielded higher values than the same plasma samples assayed after 1:4 dilution with buffer B of Hales & Randle (1963). Addition of ovine insulin (0–200μ-u./ml.) to ovine plasma showed that the sensitivity of the assay was greater in undiluted plasma than in buffer, the decrease in antibody-bound labelled insulin for a given increment in insulin concentration being greater in the plasma system. Addition of 0·01 m-EDTA (Morgan, Sorenson & Lazarow, 1964; Sheldon & Taylor, 1965) did not eliminate this discrepancy. Insulin standards in buffer did not appear to provide a suitable baseline for assay of insulin in undiluted ovine plasma.
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