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Enzyme‐Linked Immunosorbent Assay of Substance P: A Study in the Eye
Author(s) -
Stjernschantz Johan,
Gregerson Dale,
Bausher Larry,
Sears Marvin
Publication year - 1982
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1982.tb07908.x
Subject(s) - conjugate , chromatography , chemistry , alkaline phosphatase , radioimmunoassay , substance p , antibody , enzyme , absorbance , antigen , acid phosphatase , microbiology and biotechnology , biochemistry , biology , immunology , receptor , mathematics , neuropeptide , mathematical analysis
A solid phase enzyme‐linked immunosorbent assay for quantitation of substance P is presented. The assay measures the capacity of soluble substance P to compete with the solid phase antigen for a limited quantity of specific substance P antibody. The solid‐phase antigen consists of a synthetic substance P. poly‐D‐glutamic acid conjugate coated to polystyrene micro‐ELISA plate wells. Soluble substance P and antibodies to substance P are first preincubated together and then added to the wells containing solid‐phase antigen. Subsequently the wells are incubated with anti‐antibodies conjugated to alkaline phosphatase. The wells are finally incubated with p ‐nitrophenyl phosphate and the absorbance is read in a spectrophotometer 16‐24 h after the start of the assay. The threshold for detection of substance P was 5‐10 pg per well (0.25 ml). Substance P was extracted from rabbit eyes and the values obtained with the present method are compared with previously reported values based on radioimmunoassay.