An ELISA is used to recognize and quantify a specific protein in a protein mixture and is useful in protein production and purification since technicians must be able to determine the presence and concentration of a protein of interest. Two trials of and ELISA were done with 3 sub-trials of pig, sheep, and cow serum and a negative control in hopes that the pig anti-IgG antibody would recognize the pig -IgG antigen in only pig serum and not in any other serums. Samples will turn blue when exposed to TMB because the HRP bound to the anti-IgG will react. When exposed to HCl, any samples that were blue will turn yellow and one can quantify the reaction by quantifying degree of blueness and yellowness. In the first trial, HCl, which stops reaction of HRP, was added after 20 minutes when there was no reaction in any wells. This is longer than stated in the protocol. However, other trials of the same ELISA by others turned blue after 24 hours. Perhaps the TMB was exposed to light for too long before use and so had a slow
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