Arylformamidase (AFMID) is the second enzyme of the kynurenine pathway metabolizing tryptophan to nicotinic acid and nicotinamide adenine dinucleotide cofactors. Inhibition of AFMID by organophosphorus insecticides in developing chicken embryos is correlated with lowered NAD levels and severe teratogenesis.The cDNA sequence previously identified for mouse liver AFMID was cloned and expressed in Escherichia coli. Residues identified as potential catalytic triad members through sequence motif and homology modeling were mutated to alanine to probe their contributions to enzyme activity. The wild-type and mutant AFMIDs were expressed as amino terminal 6 x His-tagged recombinant proteins to facilitate purification. Three chromatography steps isolated highly purified proteins for enzyme activity comparisons.
Human Probable arylformamidase (AFMID) ELISA Kit employs a two-site sandwich ELISA to quantitate AFMID in samples. An antibody specific for AFMID has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyAFMID present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for AFMID is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of AFMID bound in the initial step. The color development is stopped and the intensity of the color is measured.
Human Probable arylformamidase (AFMID) ELISA Kit listed herein is for research use only and is not intended for use in human or clinical diagnosis. Suggested applications of our products are not recommendations to use our products in violation of any patent or as a license. We cannot be responsible for patent infringements or other violations that may occur with the use of this product.
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