Rigaudy P, Simon S, Hunter T, Sollazzo M, Billetta R, Zanetti M, Eckhart W
Molecular Biology and Virology Laboratory, Salk Institute, San Diego, CA 92186.
DNA Cell Biol. 1994 Jun;13(6):585-91. doi: 10.1089/dna.1994.13.585.
To elicit antibodies directed specifically against the neuron-specific form of the c-src gene product, pp60c-src(+), we used an antigenized antibody comprising a decamer containing the amino acid sequence specific to pp60c-src(+) inserted into the third hypervariable loop of the heavy (H)-chain variable (V)-region. This was used to raise anti-idiotype antibodies reacting with the peptide epitope in rabbits. The antisera reacted with pp60c-src(+), as judged by immune blotting, immunoprecipitation, immune complex kinase assay, and indirect immunofluorescence staining, but did not react with the fibroblast form of the c-src gene product, pp60c-src. Antigenized antibody is a useful approach for producing antibodies able to distinguish between isoforms of the same gene product and specific for the neuronal form of the Src protein.
为了诱导出特异性针对c-src基因产物的神经元特异性形式pp60c-src(+)的抗体,我们使用了一种抗原化抗体,该抗体包含一个十肽,其含有插入重链(H)可变区(V)的第三个高变环中的pp60c-src(+)特异氨基酸序列。用其在兔体内产生与肽表位反应的抗独特型抗体。通过免疫印迹、免疫沉淀、免疫复合物激酶分析和间接免疫荧光染色判断,抗血清与pp60c-src(+)反应,但不与c-src基因产物的成纤维细胞形式pp60c-src反应。抗原化抗体是产生能够区分同一基因产物的不同异构体且对Src蛋白的神经元形式具有特异性的抗体的一种有用方法。