论文部分内容阅读
cAMP抗血清是cAMP放射免疫分析的特异试剂,本文介绍其制备方法如下。 一、免疫原制备:cAMP是小分子,需与大分子载体蛋白结合后才具有免疫原性。参考Oliver等和Steiner等的方法,取牛血清白蛋白(BS-A)和血蓝蛋白(LHC)各30mg分别溶于1.5ml水中,调pH至5.5。另取ScAMP(2’-0-琥珀酰-cA-MP)15mg二份。各溶于1.5ml水中,调pH至5.5,分别滴入BSA和LHC溶液中。各加入水溶性碳二亚胺(EDC)15mg,使反应液的载体蛋白:ScAMP:EDC=2:1:1(W/W)。搅拌混匀后,暗处反应24小时,对生理盐水磷酸缓冲液(0.01M,pH7.4)在4℃透析3～7天,冰冻干燥,贮于-20℃冰箱备用。经纸电泳和紫外吸收光谱分析,证明ScAMP分别与B
The cAMP antiserum is a specific reagent for cAMP radioimmunoassay. The preparation method of the cAMP antisera is described as follows. First, the preparation of the immunogen: cAMP is a small molecule, to be immunogenicity after binding with the macromolecular carrier protein. According to the method of Oliver et al. And Steiner et al., 30 mg each of bovine serum albumin (BS-A) and hemocyanin (LHC) was dissolved in 1.5 ml of water respectively and adjusted to pH 5.5. Another take ScAMP (2’-0-succinyl-cA-MP) 15mg two copies. Each dissolved in 1.5ml of water, adjusted to pH 5.5, were dropped into BSA and LHC solution. 15 mg of water-soluble carbodiimide (EDC) was added to each reaction solution so that the carrier protein of the reaction solution: ScAMP: EDC = 2: 1: 1 (W / W). After mixing and mixing, the reaction was carried out in darkness for 24 hours. The solution was dialysed against physiological saline solution (0.01 M, pH7.4) for 3 to 7 days at 4 ° C., freeze-dried and stored in a -20 ° C. refrigerator. The paper electrophoresis and UV absorption spectroscopy showed that ScAMP and B respectively