A photoelectrochemical biosensor for detecting histone acetyltransferase activity and its preparation method
An acetyltransferase and biosensor technology, applied in electrochemical variables of materials, scientific instruments, instruments, etc., can solve the problems of operator health hazards, expensive antibody reagents, cumbersome sample processing, etc., to improve detection sensitivity and realize small instruments. The effect of simple chemical and detection methods
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[0052] (1) Preparation of catalytic reaction solution: mix 20-50 μL of different concentrations of histone acetyltransferase, 20-50 μL of substrate peptide (concentration of 10-40 μM), 10-40 μL of acetyl-CoA (concentration of 100-300 μM) well mixed. The mixture is then incubated at 20-50°C for 1-5 hours. The resulting solution was labeled as catalytic reaction solution.
[0053] (2) Preparation of exfoliated tungsten sulfide: weigh 50-300mg WS 2 And 50-300mg of polyacrylic acid dissolved in 20-60mL of water, ultrasonic 2-6h. Centrifuge the green dispersion at 2000-10000rpm for 5-30min, collect the supernatant and centrifuge at 5000-10000rpm for 5-30min, wash the obtained precipitate with deionized water several times to remove the residue, and finally dry it under vacuum.
[0054] (3) Preparation of polydopamine: Dissolve 100-200mg of dopamine hydrochloride in 50-100mL of deionized water, under vigorous stirring, take 500-1000μL of 1-3mol / L NaOH solution and add it to the a...
Embodiment 1
[0072] Example 1: Histone acetyltransferases catalyze acetylation of substrate peptides
[0073] Mix 20 μL of 50 nM histone acetyltransferase, 20 μL of substrate peptide (concentration of 20 μM), and 20 μL of acetyl-CoA (concentration of 100 μM). The mixture was then incubated at 37°C for 1 hour. The resulting solution was labeled as catalytic reaction solution.
Embodiment 2
[0074] Embodiment 2: Preparation of exfoliated tungsten sulfide
[0075] Weigh 50mg WS 2 And 300mg of polyacrylic acid dissolved in 60mL of water, ultrasonic 6h. The green dispersion was centrifuged at 3000rpm for 10min, the supernatant was collected and centrifuged at 5000rpm for 15min, the obtained precipitate was washed several times with deionized water to remove the residue, and finally dried under vacuum.
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