Renzi D, Maillet S, Grassi J, Mantellini P, Santicioli P, Theodorsson E, Surrenti C, Geppetti P
Department of Clinical Pathophysiology, University of Florence, Italy.
J Neurochem. 1991 Jan;56(1):281-6. doi: 10.1111/j.1471-4159.1991.tb02593.x.
A solid-phase enzyme immunoassay for quantitation of tachykinin-like immunoreactivity (TK-LI) is presented. Because the antiserum K-12 recognizes various tachykinins, such as neurokinin A (100%), kassinin (103%), eledoisin (51%), neurokinin B (18%), physalaemin (0.7%), and substance P (0.7%), the immunoreactivity detected in this enzyme immunoassay has been termed TK-LI. The assay was performed on 96-well microtiter plates coated with a mouse monoclonal second antibody. After preincubation of soluble neurokinin A or samples and K-12 antiserum for 3 h at room temperature, acetylcholinesterase-labelled neurokinin A was allowed to react overnight at 4 degrees C. Samples were finally incubated with Ellman's reagent for 2 h and the absorbance was measured at 414 nm. The threshold for detection of TK-LI was 2 fmol/well. TK-LI release from guinea pig dorsal spinal cord slices was evoked by capsaicin or high K+ medium. The capsaicin-evoked TK-LI release was increased in the presence of thiorphan, but not in that of captopril.
本文介绍了一种用于定量速激肽样免疫反应性（TK-LI）的固相酶免疫测定法。由于抗血清K-12能识别多种速激肽，如神经激肽A（100%）、肛褶蛙皮素（103%）、伊索雷定（51%）、神经激肽B（18%）、雨蛙肽（0.7%）和P物质（0.7%），因此在这种酶免疫测定法中检测到的免疫反应性被称为TK-LI。该测定在包被有小鼠单克隆二抗的96孔微量滴定板上进行。将可溶性神经激肽A或样品与K-12抗血清在室温下预孵育3小时后，使乙酰胆碱酯酶标记的神经激肽A在4℃下反应过夜。样品最后与埃尔曼试剂孵育2小时，并在414nm处测量吸光度。TK-LI的检测阈值为2 fmol/孔。辣椒素或高钾培养基可诱发豚鼠背脊髓切片释放TK-LI。在存在硫磷酰胺的情况下，辣椒素诱发的TK-LI释放增加，但在存在卡托普利的情况下则没有增加。
