摘要
实验建立了检测肝细胞膜低密度脂蛋白受体的抗配体抗体酶联免疫吸附测定法．测定中，固相膜受体蛋白量由包放前后膜蛋白浓度差值计算确定；低密度脂蛋白结含量按双抗体夹心法制作的低密度脂蛋白标准曲线确定，膜蛋白非村异吸附则用与酶联抗体来源相同的同种动物血浆低密度脂蛋白平行抑制试验消除．兔肝细胞膜低密度脂蛋白受体结合活性经Scatchard作图，Kd＝13．6mg／L，Bmax＝124μg／g膜蛋白（n=5）．测定结果说明建立的方法安全可靠，能反映受体的结合活性，在高脂血症及心血管病的研究中具有广泛的应用价值．本研究还对选择确定受体包被浓度、酶交联物稀释度等的棋盘滴定方法作了详细介绍和讨论．
Aim An enzyme linked immunosorbent assay (ELISA ) using HRP-conjugated rabbit antihurnan plasma apo B100 IgG was develoPed for measurement of low density liPoprotein(LDL) receptors on purified rabbit liver plasma membranes.Methods Membrane protein adhered to tbe solid phase by an overnight incubation, and was quantified by measuring the protein concentrations te fore and af-ter coating. A curve of anti-apo Bloo bo binding to known amounts of LDL was constructed to quantify the LDL bound. Parallel samples with 25-fold excess rabbit plasma LDL were assay6d to detect nonsPeCific binding.Results Kd and Bmax, worked out by scatchard plot, were l3. 59 mg/L and l24 ng/g membrane pro-tein(n= 5),re8PeCtively.C.ncluslous The results support the use of ELISA to measure LDL receptors, particularly for physiologic studies.
出处
《中国动脉硬化杂志》
CAS
CSCD
1997年第1期67-70,共4页
Chinese Journal of Arteriosclerosis
关键词
低密度脂蛋白
受体
肝细胞膜
ELISA
Receptor,low density lipoproteinl Liver plasma membrane
Enzyrne linked immunosorbent assay
