摘要
^(125)Ⅰ标记神经肽 r(NPY)做为放射配体,建立了 PY 受体放射配体平衡结合分析法。差速离心制备组织膜粗制品,膜浓度在0.02～0.2mg/ml 范围内与^(125)Ⅰ-NPY 特异结合成正比.平衡结合反应在37℃水浴条件下50分钟达平衡,最适 NpH 范围为7.0～7.4。离心法分离游离^(125)Ⅰ—INY,非特异吸附小于10%.竞争抑制试验证明^(125)Ⅰ—NPY 与膜受体结合特异性良好,非标记 NPY 的 IC50为0.44nmol/L。NPY 膜受体对胰蛋白酶和巯基抑制剂十分敏感。饱和实验及 Scatchard 作图分析表明小鼠大脑皮质 NPY 受体 Bmax=0.40～0.50pmol/mg 蛋白、kd=0.30～0.
The specific binding of^(125)INPY to membranes from mouse cerebral cortex was investigated using equilibrium binding and kinetic assay.The membranes were preparated by different—speed centrifugation.The equilibrium binding of^(125)I—NPY to membranes at 37℃ to equilibrium was by 50 minute and showed sharp optimum at PH7.0—7.4.The bound,which can't inhibited by other related peptides,was inhibited by unlabled NPY spe—cifically(IC50=0.4nmoi/L).The binding sites for^(125)I—NPY were sensitive to treatment with tripsin and thiol reagents.The
equilibrium binding of^(125)I-NPY to cerebral cortex at 37℃ was characterized by a Kd value of 0.30—0.40nmol/L and the receptor densities were 0.40—0.45 pmol/mg protein.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
1992年第4期223-226,共4页
Chinese Journal of Immunology
关键词
神经肽Y受体
放射配体分析
Neuropeptide—Y receptor Radio—ligand assay
