Protein Concentration: 20μM
- Incubate α-syn in the differential media at a concentration of 200 μM for 24 h prior to adding to the cells. The protein is very soluble at this concentration.
- After 24h, add 10 μl of this solution to the cells.
- Incubate for 24h and do the MTT assay (48h for LDH assay)
- If you want to do dose dependence with a drug, prepare the dilutions of the drug in the media in 70 μl volume (for 6 replicates). Add 70 μl of 200 μM α-syn to the drug solution. Take out 10 μl of media from the plated cells and add 20 μl of the peptide+ drug to the cells (6 replicates). Do MTT/LDH as required.
- The viability of 20μM α-syn is ~60-70% by MTT