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Variation in Enzyme‐linked lmmunosorbent Assay (ELISA) Components to Lower Sulfathiazole Detection Limits
Author(s) -
THOMSON CARRIE A.,
SPORNS PETER
Publication year - 1995
Publication title -
journal of food science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.772
H-Index - 150
eISSN - 1750-3841
pISSN - 0022-1147
DOI - 10.1111/j.1365-2621.1995.tb06250.x
Subject(s) - chemistry , horseradish peroxidase , chromatography , conjugate , detection limit , reagent , enzyme , biochemistry , organic chemistry , mathematical analysis , mathematics
Some basic components of an indirect competitive enzyme‐linked immunosorbent assay (ELISA) were manipulated to reduce sulfathiazole (ST) detection limits. 3.3′.5.5′‐Tetramethvlbenzidine (TMB) was more kffeitive than several aliemative chromogens tested for the hetection of horseradish peroxidase activity. Color development was maximized at pH 4.0. Using TMB and a 2M sulfuric acid stopping reagent, an assay was develoued using highly diluted (1:500,000) rabbit serum and low levels (0.0313 μg/mY.), of coating conjugate, with a C value (inflection point of sigmoid standard curve) of 4.4 ppb ST. Urease was unsuitable as a marker for indirect competitive ELISAs.