ADAM15 Protein, Mouse (HEK293, His)

Materials
ADAM15 Protein, Mouse (HEK293, His) (HY-P75564)
Cell Line for Activity Assay: Jurkat Cells
Cell Culture Medium: 1640 Medium + 10% FBS + 1% P/S
Penicillin-Streptomycin (100×), Sterile (HY-K1006)
Cell Counting Kit-8 (CCK8 Solution, HY-K0301)

Procedure
1. Dilute Mouse ADAM15 protein to various concentrations with PBS and add 40 μL to each well of a 96-well plate, making the final concentrations of Mouse ADAM15 protein in each well 0, 0.01, 0.05, 0.1, 0.5, 1, 2, 3.9, 7.8, 15.6, 31.25, 100, 600 μg/mL.
2. Place the 96-well plate in a ventilated and dry place to dry for 1 h, then store at 4°C overnight.
3. Add 50 μL of 1 mg/mL BSA solution to each well and block at 37°C for 1 h.
4. Discard the BSA and wash the plate twice with serum-free 1640 medium, 200 μL each time.
5. Seed cells into the coated 96-well plate at a density of 100,000 cells/well, with a volume of 100 μL per well.
6. Incubate the cells in a 37°C, 5% CO₂ incubator for 4 h.
7. Discard the medium in the wells and wash twice with PBS.
8. Under dark conditions, add 100 μL of 10% CCK8 solution to each well and incubate overnight in a 37°C, 5% CO₂ incubator.
9. Measure the OD value at 450 nm using a microplate reader.
10. Use GraphPad Prism software to plot a curve with OD value as the ordinate and protein concentration as the abscissa, and calculate the ED₅₀ value.