中文摘要：

探讨两种新的检测方法—In-cell Western和Cell ELISA在大分子量蛋白质相对定量中的应用。将培养细胞原位固定、通透化、封闭,然后加入目标蛋白质的一抗、红外荧光标记的二抗或酶标记的二抗,使用双红外成像系统或多功能酶标仪对目标蛋白质进行相对定量。通过这两种方法检测模拟失重环境对成骨细胞中大分子量骨架相关蛋白——微管微丝交联因子（microtubule actin cross-linking factor 1,MACF1）含量的影响,结果表明模拟失重环境促进了MACF1表达,且这两种方法检测结果一致。In-cell Western和Cell ELISA这两种检测方法灵敏迅速、操作简便,可以用于大分子量蛋白质的相对定量。

英文摘要：

The purpose of this study is to discuss two new methods, In-cell Western and Cell ELISA, which are used to relatively quantify the large molecular weight proteins. Cultured cells were fixed in situ, permeabUzed, blocked, and then primary antibody for the aim protein and secondary antibody labeled by IRDye^TM 800 or horseradish peroxidase （HRP） were added respectively. The aim proteins were relatively quantified using Infrared Imaging System or Multi-Mode Microplate Reader. The expression of microtubule actin cross-linking factor 1 （MACF1）, a cytoskeleton related protein of large molecular weight, was detected by the two methods in osteoblast. The results of both In-cell Western and Cell ELISA showed that the expression of MACF1 was increased under simulated weightlessness environment. With the advantages of sensitivity, rapidity, and convenience, In-cell Western and Cell ELISA are therefore useful two new methods to relatively quantify the large molecular weight proteins.