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I am using undiluted serum and plasma samples and observing high OD values. Do you have any suggestion why is this so?
Asked on Sep 01 2016
This is most likely due to matrix effect which affect measurement of analyte of interest in ELISA when undiluted serum or plasma samples are used. Matrix effect can arise from a number of matrix components such as phospholipids, carbohydrates, and endogenous metabolites (bilirubin), residual formulation components from intraperitoneal (ip), intravenous (iv), or oral dosing (po) experiments; for example, polyethylene glycol (PEG), solutol, polysorbate (Tween 80), etc, an interaction between the analyte of interest and the matrix, such as covalent binding to plasma proteins or the enzymatic degradation of a prodrug. The easiest way to overcome this is by using diluted samples e.g. 1/2 or 1/10 dilution.
Answered on Sep 01 2016