Wilder R L, Yuen C C, Mage R G
J Immunol. 1979 Feb;122(2):459-63.
Rabbit and mouse splenic lymphocytes were radioiodinated by the lactoperoxidase technique, extracted with non-ionic detergent, immunoprecipitated with high titered rabbit anti-kappa antisera, and compared by SDS-PAGE. Mouse sIg peaks were reproducibly larger in size than rabbit sIg peaks (often greater than 10 times). Neither differences in incorporation of label into the rabbit cell surface, nor differences in average sIg density explain this result. Total TCA-precipitable radioactivity was similar in each species. Estimation of the relative amounts of sIg in the mouse and rabbit showed similar average sIg densities. Differences in detergent solubility, proteolytic lability, or antisera used also do not adequately account for this difference. Thus, these data indicate that radioactivity incorporated after lactoperoxidase catalyzed cell surface radioiodination may not reflect cell surface Ig density. Conclusions about cell surface density based upon relative incorporation of radioactivity should be confirmed by other approaches.
兔和小鼠的脾淋巴细胞通过乳过氧化物酶技术进行放射性碘化,用非离子去污剂提取,用高滴度兔抗κ抗血清进行免疫沉淀,并通过SDS-PAGE进行比较。小鼠sIg峰的大小比兔sIg峰可重复性地更大(通常大于10倍)。无论是标记物掺入兔细胞表面的差异,还是平均sIg密度的差异,都无法解释这一结果。每个物种中总的三氯乙酸可沉淀放射性相似。对小鼠和兔中sIg相对量的估计显示平均sIg密度相似。去污剂溶解性、蛋白水解稳定性或所用抗血清的差异也不能充分解释这种差异。因此,这些数据表明,乳过氧化物酶催化细胞表面放射性碘化后掺入的放射性可能无法反映细胞表面Ig密度。基于放射性相对掺入量得出的关于细胞表面密度的结论应通过其他方法进行证实。