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尿中上皮成长因子(hEGF)由53种氨基酸组成,尿中含量25～250μg/ml,留置标本方便,临床价值较高。放免测定(RIA):准备:1.标准 hEGF 可用氯胺T 法制作。即 hEGF(2μg/20μl),0.5M 磷酸缓冲液(pH7.5),Na~(125)Ⅰ(1mci/10μl),氯胺 T(25μg/10ml),室温中反应20秒钟,加焦氯亚硫酸盐(60μg/10μl)至反应停止。加含2%苯磺酸(BSA)磷酸缓冲液,G25号和 G50号凝胶,过滤即得到精制标准~(125)IhEGF。其活性为150～300μci/μg。2.缓冲液:50mM(毫摩尔)Na_2HPO_4-KH_2PO_4,77 mMNacl,0.5%BSA,pH7.4。操作:hEGF 抗体(1:40,000)200μl及标准 hEGF 及稀释尿标本100μl,~(125)IhEGF(～10~4cpm)100μl,在4℃、放置48小时。然后加入正常家兔血清(1:100)100μl、抗家兔 IgG 山羊血清(1:20)100μl,在4℃、放置1夜,低温离心、(4℃、3000转、30分钟),吸取上清液测定沉渣的放射活性。此法是
Urine epithelial growth factor (hEGF) consists of 53 kinds of amino acids, urine content of 25 ~ 250μg / ml, indwelling specimens convenient, high clinical value. Radioimmunoassay (RIA): Preparation: 1. Standard hEGF available chloramine T method. Namely, hEGF (2μg / 20μl), 0.5M phosphate buffer (pH7.5), Na125I (1mci / 10μl) and chloramine T (25μg / 10ml) Sulfite (60 μg / 10 μl) until the reaction is stopped. Add 2% benzenesulfonic acid (BSA) phosphate buffer, G25 and G50 gels and filter to give purified standard 125 IhEGF. Its activity is 150 ~ 300μci / μg. 2. Buffer: 50 mM (millimoles) Na 2 HPO 4 -KH 2 PO 4, 77 mM NaCl, 0.5% BSA, pH 7.4. Operation: 200 μl of hEGF antibody (1: 40,000), 100 μl of standard hEGF and diluted urine samples, and 100 μl of ~ (125) IhEGF (~10 ~ 4 cpm) were added and the plate was allowed to stand at 4 ° C for 48 hours. Then, 100 μl of normal rabbit serum (1: 100) and 100 μl of anti-rabbit IgG goat serum (1:20) were added and the plate was allowed to stand overnight at 4 ° C and centrifuged at 3000 rpm for 30 minutes at 4 ° C Serum was measured radioactive sediment. This method is