Ducommun B, Beach D
Cold Spring Harbor Laboratory, New York 11724.
Anal Biochem. 1990 May 15;187(1):94-7. doi: 10.1016/0003-2697(90)90422-6.
A microassay for p34cdc2 based on the high affinity association between cdc2 and Schizosaccharomyces pombe p13suc1 has been developed. p13 purified from Escherichia coli was immobilized on microtiter plates and cellular lysate was incubated in the wells to allow the binding of cdc2 and its associated proteins. p34cdc2 was assayed either as a histone kinase or by immunological methods. The method was optimized for S. pombe cell extracts but can also be applied to other organisms such as Xenopus oocytes or HeLa cells. This rapid assay allows the specific determination of p34cdc2 histone H1 kinase activity in a very large number of samples.
基于cdc2与粟酒裂殖酵母p13suc1之间的高亲和力结合，已开发出一种用于p34cdc2的微量测定法。从大肠杆菌中纯化的p13固定在微量滴定板上，细胞裂解物在孔中孵育，以使cdc2及其相关蛋白结合。p34cdc2可作为组蛋白激酶进行测定，也可通过免疫学方法进行测定。该方法针对粟酒裂殖酵母细胞提取物进行了优化，但也可应用于其他生物体，如非洲爪蟾卵母细胞或HeLa细胞。这种快速测定法能够在大量样品中特异性地测定p34cdc2组蛋白H1激酶活性。
