Application of flavonoids
A technology of flavonoids and compounds, applied in the application field of flavonoids, can solve the problems of cytotoxicity, low blood drug concentration, narrow anti-tumor spectrum, etc.
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[0059] Substrate peptide STTPGGTLFSTTPG (the sequence of the substrate peptide is shown in SEQIDNO: 1 in the sequence listing, referring to the mTOR kinase test method of PerkinElmer Company), the substrate peptide was synthesized by Jill Biochemical (Shanghai) Co., Ltd., and its N 5-carboxyfluorescein (5-FAM) labeling is carried out at the end to facilitate fluorescence detection and improve the sensitivity of detection. The peptide structure of the obtained substrate peptide is shown in formula II:
[0060]
[0061]
[0062] The mTORC1 used in the examples was purchased from Sigma Reagent Co., Ltd., product number: SRP0364.
[0063] Prepare 25 mL of enzyme reaction buffer: 50 mM 4-hydroxyethylpiperazineethanesulfonic acid (Hepes), pH adjusted to 7.5 by 4M NaOH, 2 mM 1,4-dithiothreitol, 10 mM magnesium chloride, 1 mM ethylene glycol bis(2 -aminoethyl ether) tetraacetic acid, 0.01% (v / v, the same below) Tween 20 and 3mM manganese chloride.
[0064] Use 2 μL of enzyme re...
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