High-throughput screening system and screening method for potential drugs targeting c-cbl protein ring functional domain
A technology of c-cbl and functional domains, which is applied in the field of high-throughput screening of potential drugs targeting the circular functional domain of c-Cbl protein, achieving good application prospects, low cost and high accuracy
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0035] Example 1 Screening of Potential Drugs Targeting the c-Cbl Ring Domain Protein (c-Cbl Ring Domain Protein Ligand)
[0036]A mixture of FITC-P1 at a final concentration of 100 nM and a c-Cbl ring domain protein at a final concentration of 2 μM was prepared, and then this mixture was added to a 384-well plate at 19 μL / well. The unknown compound was dissolved in 20 μL DMSO to prepare a 200 μM stock solution, and the stock solution was diluted 7 times in half to obtain 8 different concentrations of the samples to be tested (the concentration range was 200 μM-1.56 μM). Add 1 μL / well of the sample to be tested into a 384-well plate (final concentration range 10 μM-0.08 μM). Shake the 384-well plate for 30 seconds before the test to fully mix the solution, and use the ZS-2 plate reader to read the polarized fluorescence value of the sample. The instrument should be set at Ex554 / Em575. Each concentration of each sample should be tested 3 times.
[0037] After the instrument ...
Embodiment 2
[0038] Example 2 Screening of potential drugs (c-Cbl ring domain protein ligands) targeting c-Cbl ring domain protein
[0039] A mixture of FITC-P2 at a final concentration of 100 nM and a c-Cbl ring domain protein at a final concentration of 2 μM was prepared, and then this mixture was added to a 384-well plate at 19 μL / well. The unknown compound was dissolved in 20 μL DMSO to prepare a 200 μM stock solution, and the stock solution was diluted 7 times in half to obtain 8 different concentrations of the samples to be tested (the concentration range was 200 μM-1.56 μM). Add 1 μL / well of the sample to be tested into a 384-well plate (final concentration range 10 μM-0.08 μM). Shake the 384-well plate for 30 seconds before the test to fully mix the solution, and use the ZS-2 plate reader to read the polarized fluorescence value of the sample. The instrument should be set at Ex554 / Em575. Each concentration of each sample should be tested 3 times.
[0040] After the instrument re...
Embodiment 3
[0041] Example 3 Screening of Potential Drugs Targeting the c-Cbl Ring Domain Protein (c-Cbl Ring Domain Protein Ligand)
[0042] A mixture of FITC-P3 at a final concentration of 100 nM and a c-Cbl ring domain protein at a final concentration of 2 μM was prepared, and then this mixture was added to a 384-well plate at 19 μL / well. The unknown compound was dissolved in 20 μL DMSO to prepare a 200 μM stock solution, and the stock solution was diluted 7 times in half to obtain 8 different concentrations of the samples to be tested (the concentration range was 200 μM-1.56 μM). Add 1 μL / well of the sample to be tested into a 384-well plate (final concentration range 10 μM-0.08 μM). Shake the 384-well plate for 30 seconds before the test to fully mix the solution, and use the ZS-2 plate reader to read the polarized fluorescence value of the sample. The instrument should be set at Ex554 / Em575. Each concentration of each sample should be tested 3 times.
[0043] After the instrument...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More