Holmes E, Engvall E
La Jolla Cancer Research Foundation, California 92037.
Anal Biochem. 1993 Oct;214(1):100-5. doi: 10.1006/abio.1993.1462.
An assay for the determination of relative concentrations of integrins on clonal cultured cells is described. Unpurified monoclonal antibodies to integrin subunits, present in ascites or hybridoma culture media, are immobilized on a plastic surface via goat antibodies to mouse IgG. Cell attachment to the integrin-coated substrate is quantitated after fixation and staining of the cells. The assay is specific and very sensitive; only cells with the relevant integrins attach to antibody-coated substrates and concentrations of antibody as low as 1-10 ng/ml are sufficient. Titrations of the antibodies on the solid phase allow the estimation of the relative amounts of integrins on different cell types. The results with this assay correlate well with results obtained by flow cytometry.
本文描述了一种用于测定克隆培养细胞上整合素相对浓度的检测方法。存在于腹水或杂交瘤培养基中的针对整合素亚基的未纯化单克隆抗体,通过抗小鼠IgG的山羊抗体固定在塑料表面。在细胞固定和染色后,对细胞与整合素包被底物的附着情况进行定量。该检测方法具有特异性且非常灵敏;只有带有相关整合素的细胞才会附着在抗体包被的底物上,低至1 - 10 ng/ml的抗体浓度就足够了。对固相上抗体进行滴定可估计不同细胞类型上整合素的相对含量。该检测方法的结果与通过流式细胞术获得的结果相关性良好。