A biosensing assay method for detecting estrogen-binding activity in water samples
A technology combining activity and estrogen, which is applied in the direction of material stimulation analysis, material analysis, biological testing, etc., can solve the problems of increasing detection complexity and detection cost, inability to fully characterize estrogen binding activity, etc., and achieve easy online and in situ The effect of monitoring, reducing detection costs, and stabilizing regeneration
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[0031] 1) Make a receptor protein affinity column
[0032] Resuscitate the Escherichia coli transformed with the His-ER expression plasmid, smear it on the LB plate containing 50 mg / L kanamycin, pick a single clone into the LB medium containing kanamycin after 16 hours, and culture with shaking at 37°C At 20 hours, absorb 15mL to 100mL LB medium containing 15% sucrose, culture at 37°C for 3h with shaking, then cool down gradually, and the temperature drops to 15°C within 3h, and add IPTG with a final concentration of 0.5mM for low temperature induction for 16h. Centrifuge the bacterial liquid to obtain the bacterial cells, add 10 mL of lysis buffer to resuspend the bacterial cells, crush on an ultrasonic breaker, and centrifuge at a high speed and low temperature to obtain the supernatant. Add the supernatant to a 200 μL nickel column agarose gel column, shake and incubate at 4°C for 2 hours, centrifuge to discard the supernatant, wash the impurity protein with purification bu...
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