Method for detecting environmental estrogen interferent in environmental pollution
A technology for environmental estrogen and environmental pollutants, applied in the field of pollutant detection, can solve problems such as incompleteness, and achieve the effect of improving detection efficiency and increasing experimental throughput
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Embodiment 1
[0057] Embodiment 1, the detection of environmental pollutant bisphenol A (BPA)
[0058] 1. Cell culture
[0059] Human breast cancer cell line SKBR3 cells were cultured at 37°C, 5% (v / v) CO 2 In the cell culture incubator, the cell culture medium of SKBR3 is containing 10% (v / v) fetal bovine serum (Fetal bovine serum, FBS, Gibco) and 1% (w / v) penicillin-streptomycin (penicillin / streptomycin, Gibco) RPMI 1640 medium.
[0060] During the cell culture process, the cell culture medium was replaced every 2d-3d. When the confluence of the cells reached more than 85%, the cells were digested with 0.05% (w / v) trypsin (Trypsin-EDTA, Gibco), and the passaged cell density was adjusted according to the needs of the experiment. .
[0061] 2. Exposure of environmental pollutants in 96-well plate
[0062] (1) Cell incubation
[0063] SKBR3 cells in 1.5 × 10 4 The density per well was plated in 96-well plates, and after 24 hours of incubation in complete medium with phenol red, the log...
Embodiment 2
[0075] Embodiment 2, the detection of different drinking water samples
[0076] 1. Cell culture
[0077] Human breast cancer cell line SKBR3 cells were cultured at 37°C, 5% CO 2 In the cell culture incubator, the cell culture medium of SKBR3 was RPMI 1640 medium containing 10% fetal bovine serum (Fetal bovine serum, FBS, Gibco) and 1% penicillin / streptomycin (Gibco).
[0078] During the cell culture process, the cell culture medium was replaced every 2d-3d. When the confluence of the cells reached more than 85%, the cells were digested with 0.05% trypsin (Trypsin-EDTA, Gibco), and the passaged cell density was adjusted according to the experimental needs.
[0079] 2. Exposure of environmental pollutants in 96-well plate
[0080] (1) Cell incubation
[0081] SKBR3 cells were 2 × 10 4 The density per well was plated in 96-well plates, and after 24 hours of incubation in complete medium with phenol red, the logarithmic phase was reached.
[0082] (2) test
[0083] 1) Replace ...
Embodiment 3
[0091] Embodiment 3, the detection of environmental pollutant bisphenol A (BPA)
[0092] 1. Cell culture
[0093] SKBR3 cells were cultured at 37°C, 5% (v / v) CO 2 In the cell culture incubator, the cell culture medium of SKBR3 is containing 10% (v / v) fetal bovine serum (Fetal bovine serum, FBS, Gibco) and 1% (w / v) penicillin-streptomycin (penicillin / streptomycin, Gibco) RPMI 1640 medium.
[0094] During the cell culture process, the cell culture medium was replaced every 2d-3d. When the confluence of the cells reached more than 85%, the cells were digested with 0.05% (w / v) trypsin (Trypsin-EDTA, Gibco), and the passaged cell density was adjusted according to the needs of the experiment. .
[0095] 2. Exposure of environmental pollutants in 96-well plate
[0096] (1) Cell incubation
[0097] SKBR3 cells at 1.0 × 10 4 The density per well was plated in 96-well plates, and after 24 hours of incubation in complete medium with phenol red, the logarithmic phase was reached.
...
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