A NEW SOLID-STATE REAGENT TO IODINATE PROTEINS .1. CONDITIONS FOR THE EFFICIENT LABELING OF ANTISERUM
A NEW SOLID-STATE REAGENT TO IODINATE PROTEINS .1. CONDITIONS FOR THE EFFICIENT LABELING OF ANTISERUM
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DOI：
10.1016/0003-2697(82)90025-2
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发表时间：
1982-01-01
影响因子：
2.9
通讯作者：
MARKWELL, MAK
中科院分区：
文献类型：
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作者：
MARKWELL, MAK
用一种新的多肽和蛋白质碘化固体试剂标记抗猪胰岛素抗血清。优化pH、温度、缓冲液、反应物浓度和反应容器类型的反应条件，以实现放射性碘的几乎定量掺入（99%）、蛋白质的良好回收（92%）和中等比活性（9.9 μ Ci/μ g抗血清）。第二组条件产生高比活性（161 μ Ci/μ g）的抗血清，其保留了识别胰岛素的免疫学能力。研究了实际碘化物种的性质以及叠氮化钠、洗涤剂和离液试剂（如尿素和高盐）对反应的影响。
A new solid-state reagent for iodinating polypeptides and proteins was used to radiolabel antiporcine insulin antiserum. Reaction conditions of pH, temperature, buffer, reactant concentration, and the type of reaction vessel were optimized to achieve almost quantitative incorporation of radioiodide (99%), good recovery of protein (92%), and a moderate specific activity (9.9 .mu.Ci/.mu.g of antiserum). A second set of conditions produced antiserum of high specific activity (161 .mu.Ci/.mu.g) which retained its immunological ability to recognize insulin. The nature of the actual iodinating species and the effects on the reaction of sodium azide, detergents, and chaotropic reagents such as urea and high salt were investigated.