METHOD FOR IMMUNOFLUORESCENCE ASSAY OF EXCISION REPAIR PROTEIN ERCC1 IN HUMAN SOLID TUMOURS

摘要

FIELD: medicine.;SUBSTANCE: method for immunofluoresence assay of ERCC1 excision repair protein in human solid tumours involves preparing one-cell suspensions of tumour tissue and a reference culture, fixing in 4% formaldehyde, incubating with Tween 20, washing in phosphate buffer pH 7.4, incubating with primary (clone 8F1) and secondary (fluorescent dye DyLight 650) antibodies, washing twice in phosphate buffer pH 7.4, analysing the findings in a flow cytofluorimeter; using a monolayer culture of human breast cancer cell line MCF-7 as a reference culture; calculating a protein expression level by means of Kolmogorov-Smirnov statistical test, and calculating an expression intensity by means of a geometric mean of tumour cell fluorescence.;EFFECT: using the method enables the quantitative assessment of ERCC1 protein expression that eliminates a subjective component from the result assessment.;2 dwg