Description
Introduction:
Erythropoietin (EPO) is a heavily glycosylated protein with a molecular weight of about 30,000 – 34,000 Daltons. Rat EPO is a polypeptide consisting of 166 amino acids, containing one O-linked and three N-linked carbohydrate chains. The recombinant EPO is a good substitute for the native protein for use in an immunoassay.
Intended Use:
The EPO ELISA is intended for the quantitative determination of Erythropoietin (EPO) in rat serum.
Principle:
The method employs the quantitative sandwich enzyme immunoassay technique. Samples and standards are pipetted into microwell precoated with anti-EPO antibody . After incubation and washing, anti-EPO antibody linked to Biotin is added to the wells. EPO present in the sample and standards will form a complex in the wells. HRP Conjugate is pipetted and incubated. Washing is done to remove the unbound HRP Conjugate. The ready to use substrate solution (TMB) is added to microwells and color develops directly proportional to the amount of EPO in the standards and sample. Color development is then stopped by addition of stop solution. Absorbance is measured at 450 nm.
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