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以2,4-二硝基氟苯修饰铁传递蛋白(Tf)及小牛血清白蛋白,并制得高效价的抗2,4-二硝基苯基团(DNP)血清。运用放射受体分析法比较Tf经DNP化修饰前后与细胞膜上铁传递蛋白受体(TR)的亲和力,发现两者的亲和常数及受体总数基本一致。应用DNP-Tf,抗DNP血清为第一抗体的间接酶标检测细胞膜上的TR,结果表明该法的灵敏度是应用Tf/抗Tf血清的间接酶标法的4倍左右,并明显降低非特异性吸附。
The transferrin (Tf) and bovine serum albumin were modified with 2,4-dinitrofluorobenzene and high titer anti-2,4-dinitrophenyl group (DNP) serum was obtained. Using radioimmunoassay to compare the affinity of Tf with the transferrin receptor (TR) on the cell membrane before and after DNP modification, we found that the affinity constants and the total number of receptors were basically the same. Using DNP-Tf and anti-DNP serum as the primary antibody to detect the TR on the cell membrane, the sensitivity of this method was about 4 times higher than indirect enzyme-labeling method using Tf / anti-Tf serum and significantly reduced nonspecificity Adsorption.