Enzyme‐Linked Immunosorbent Assay of Substance P: A Study in the Eye

Johan Stjernschantz, Dale Gregerson, Larry Bausher, Marvin Sears

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

Abstract: 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.

Original languageEnglish (US)
Pages (from-to)1323-1328
Number of pages6
JournalJournal of Neurochemistry
Volume38
Issue number5
DOIs
StatePublished - May 1982

Keywords

  • Enzyme
  • Eye
  • Rabbit
  • Substance P
  • linked immunosorbent assay

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