Blood Research Institute, Versiti, Milwaukee, WI, USA.
Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, USA.
Methods Mol Biol. 2021;2217:3-15. doi: 10.1007/978-1-0716-0962-0_1.
Soluble ligand and conformation-dependent antibody binding assay of integrins on the cell surface is an effective approach to evaluate the activation status of integrins in live cells. The ligands or antibodies are usually labeled with biotin or a fluorescent dye and incubated with integrin-expressing cells in suspension. The cell-bound ligands and antibodies are then detected by flow cytometry. Here we describe the detailed protocols of soluble ligand or antibody binding assay for αβ, αβ, αβ, and αβ integrins that are transiently or stably expressed in the model cell lines such as HEK293 or CHO-k1 cells.
在活细胞中评估整合素的激活状态的有效方法是检测细胞表面整合素的可溶性配体和构象依赖性抗体结合。配体或抗体通常用生物素或荧光染料标记，并与悬浮培养的整合素表达细胞孵育。然后通过流式细胞术检测细胞结合的配体和抗体。本实验描述了瞬时或稳定表达于模型细胞系如 HEK293 或 CHO-k1 细胞中的αβ、αβ、αβ 和αβ 整合素的可溶性配体或抗体结合分析的详细方案。
