Varenne S, Piovant M, Pagès J M, Lazdunski C
Eur J Biochem. 1978 May 16;86(2):603-6. doi: 10.1111/j.1432-1033.1978.tb12344.x.
Polysomes containing nascent chains of alkaline phosphatase have been isolated from a membrane-bound polysome preparation. Indirect immunoprecipitation using conformation-specific antibodies has been employed. This technique provides a good enrichment of these polyribosomes since routinely no more than than 10--15% of non-specific immunoprecipitation was observed. The yield of the procedure is generally 40% but can be increased if higher non-specific immunoprecipitation is tolerated. Antibodies, previously described, directed against uncoiled or folded monomers of alkaline phosphatase can be used as primary antibody to recognize the nascent chains contained in membrane-bound polysomes which suggests that these chains are partially folded.
已从膜结合多聚核糖体制剂中分离出含有碱性磷酸酶新生链的多核糖体。采用了使用构象特异性抗体的间接免疫沉淀法。由于常规观察到非特异性免疫沉淀不超过10%-15%，该技术能很好地富集这些多核糖体。该方法的产量一般为40%，但如果能容忍更高的非特异性免疫沉淀，产量可以提高。先前描述的针对碱性磷酸酶未卷曲或折叠单体的抗体可作为一抗，用于识别膜结合多核糖体中包含的新生链，这表明这些链是部分折叠的。
