Voigt Wieland
Klinik für Innere Medizin IV-Martin Luther Universitat Halle, Halle/Saale, Germany.
Methods Mol Med. 2005;110:39-48. doi: 10.1385/1-59259-869-2:039.
The sulforhodamine B (SRB) assay was developed by Skehan and colleagues to measure drug-induced cytotoxicity and cell proliferation for large-scale drug-screening applications. Its principle is based on the ability of the protein dye sulforhodamine B to bind electrostatically and pH dependent on protein basic amino acid residues of trichloroacetic acid-fixed cells. Under mild acidic conditions it binds to and under mild basic conditions it can be extracted from cells and solubilized for measurement. Results of the SRB assay were linear with cell number and cellular protein measured at cellular densities ranging from 1 to 200% of confluence. Its sensitivity is comparable with that of several fluorescence assays and superior to that of Lowry or Bradford. The signal-to-noise ratio is favorable and the resolution is 1000-2000 cells/well. It performed similarly compared to other cytotoxicity assays such as MTT or clonogenic assay. The SRB assay possesses a colorimetric end point and is nondestructive and indefinitely stable. These practical advances make the SRB assay an appropriate and sensitive assay to measure drug-induced cytotoxicity even at large-scale application.
磺酰罗丹明B（SRB）测定法由斯凯汉及其同事开发，用于大规模药物筛选应用中测量药物诱导的细胞毒性和细胞增殖。其原理基于蛋白质染料磺酰罗丹明B与三氯乙酸固定细胞的蛋白质碱性氨基酸残基静电结合且依赖于pH值的能力。在轻度酸性条件下它与之结合，在轻度碱性条件下它可从细胞中提取并溶解以进行测量。SRB测定法的结果与细胞数量呈线性关系，并且在细胞密度为汇合度的1%至200%范围内测量的细胞蛋白质也呈线性关系。其灵敏度与几种荧光测定法相当，且优于洛瑞法或布拉德福德法。信噪比良好，分辨率为每孔1000 - 2000个细胞。与其他细胞毒性测定法如MTT或克隆形成测定法相比，它表现相似。SRB测定法具有比色终点，且无损且稳定性无限期。这些实际优势使得SRB测定法即使在大规模应用中也是一种合适且灵敏的用于测量药物诱导细胞毒性的测定法。
