Cancer promotion activity quantitative determination method and screening method of cancer promoter
A quantitative detection and activity technology, applied in the field of biomedicine, can solve the problems of prolonged detection time, time-consuming and laborious, unable to meet the actual detection of fast and large samples, etc., and achieve the effect of quantitative detection.
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[0062] 1. Establish a stable fluorescent probe-labeled cell model
[0063] 1.1 Cell culture
[0064] Select human immortalized epidermal HaCat cells (CCTCC, China Type Culture Collection Center), use MEM medium containing 10% fetal bovine serum and penicillin-streptomycin double antibody (penicillin 100U / mL; streptomycin 100 μg / mL), At 37°C, 5% CO 2 cultivated under conditions. The cells in the logarithmic growth phase were digested with 0.25% trypsin (containing 0.03% EDTA), and passaged once every 2-3 days.
[0065] 1.2 Fluorescent probe preparation
[0066] Calcein AM (C-3099, Molecular Probes) solution (1 mM) was aliquoted and stored at -20°C in the dark for future use. DiI (D-282, Molecular Probes) powder was prepared into 2mM stock solution with DMSO, and stored at -20°C in the dark for future use after aliquoting.
[0067] Probe diluent: Glucose was dissolved in sterilized double distilled water to prepare a 0.3M glucose use solution, and stored at 4°C for later us...
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