Abstract
An Enzyme Linked Immunosorbent Assay (ELISA) is described for use in the determination of β-endorphin antibody titers, as well as for the quantitation of naturally occuring levels of β-endorphin in plasma and other bodily fluids. The ability of the assay to accomodate unpurified samples containing small concentrations of β-endorphin was improved through the use of affinity purified antibodies in conjunction with a competitive inhibition ELISA. The problem of non-specific binding of β-endorphin during competitive inhibition assays was circumvented through a two-step process in which the plate was first coated with BSA, followed by a second plate coating with poly-lysine (MW4000). The second coating with poly-lysine was found necessary in order to eliminate intermolecular void spaces following initial plate treatment with BSA. Following these procedures enabled quantitation of β-EP at a level as low as 10pmoles per microtitre plate well.
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