Stjernschantz J, Gregerson D, Bausher L, Sears M
J Neurochem. 1982 May;38(5):1323-8. doi: 10.1111/j.1471-4159.1982.tb07908.x.
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 an the absorbance is read in a spectrophotometer 16--24 hr 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.
本文介绍了一种用于定量P物质的固相酶联免疫吸附测定法。该测定法测量可溶性P物质与固相抗原竞争有限量特异性P物质抗体的能力。固相抗原由包被在聚苯乙烯微量酶联免疫吸附测定板孔上的合成P物质-聚-D-谷氨酸结合物组成。可溶性P物质和P物质抗体首先一起预孵育，然后加入含有固相抗原的孔中。随后，孔与结合碱性磷酸酶的抗抗体一起孵育。孔最后与对硝基苯磷酸孵育，并在测定开始后16 - 24小时在分光光度计中读取吸光度。P物质的检测阈值为每孔（0.25毫升）5 - 10皮克。从兔眼中提取P物质，并将本方法获得的值与先前基于放射免疫测定报告的值进行比较。
