Protein Concentration: 15μM
Cells: PC12 cells
- Incubate calcitonin in MiliQ water at a concentration of 1.5 mM for 24 h prior to adding to the cells. The protein is very soluble at this concentration.
- Dilute this solution to 150 μM in differential media after 24h and add 10 μl of this solution to the cells.
- Incubate for 24h and do the MTT 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 150 μM calcitonin 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 15μM calcitonin is ~70-80% by MTT assay