用于特定酶周转研究的抗血清的吸收
Absorption of antisera for studies on specific enzyme turnover.
作者信息
Walker J H, Betts S A, Manning R, Mayer R J
出版信息
Biochem J. 1976 Nov;159(2):355-62. doi: 10.1042/bj1590355.
Abstract
Antisera were raised to acetyl-CoA carboxylase and 6-phosphogluconate dehydrogenase from mammary glands of lactating rabbits, and cytochrome oxidase from rat liver. The enzymes were all highly purified but gave rise to multispecific antisera when tested against tissue extracts. Absorption procedures were devised to free the antisera of contaminating antibodies. Antisera to acetyl-CoA carboxylase and cytochrome oxidase were absorbed with fractions discarded during enzyme purification. The antiserum to 6-phospho-gluconate dehydrogenase was absorbed with a tissue extract from an early stage in mammary-gland differentiation. Monospecific antisera are essential for enzyme turnover studies and therefore antisera should be extensively tested and absorbed before use. A general procedure for the absorption of antisera to purified enzymes has been devised on the basis of accepted principles of antisera absorption.
摘要
制备了针对泌乳兔乳腺中乙酰辅酶A羧化酶和6-磷酸葡萄糖酸脱氢酶以及大鼠肝脏细胞色素氧化酶的抗血清。这些酶均经过高度纯化，但在与组织提取物进行检测时会产生多特异性抗血清。设计了吸收程序以去除抗血清中污染的抗体。乙酰辅酶A羧化酶和细胞色素氧化酶的抗血清用酶纯化过程中丢弃的组分进行吸收。6-磷酸葡萄糖酸脱氢酶的抗血清用乳腺分化早期的组织提取物进行吸收。单特异性抗血清对于酶周转研究至关重要，因此抗血清在使用前应进行广泛检测和吸收。基于公认的抗血清吸收原理，设计了一种用于纯化酶抗血清吸收的通用程序。
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