FIELD: medicine.
SUBSTANCE: method involves preparation of [32P]-marked DNA-duplex, containing an AP-site. The prepared DNA-duplex is added to a reaction mixture containing analysed proteins of human cell extract and a buffer solution. The mixture is incubated during 10-15 min at temperature 35-37°C; NaBH4 is added to concentration 20-30 mM, and the ice reduction reaction is carried out within 30-60 minutes. Further, electrophoresis is used to separated proteins. Gels are exposed with an X-ray film to detect the presence of Ku-antigen in the analysed human cells by a strip observed on a gel radioautograph after electrophoresis and showing apparent molecular weight about 95 kDa, corresponding to a covalent addition product of the DNA-duplex containing the AP-site, to a Ku80-subunit of the Ku-antigen.
EFFECT: method allows for high-sensitive detection of Ku-antigen presence in the cells of various origins.
2 cl, 1 dwg, 2 ex
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