For in vivo experiments (the concentration of 1 mg/kg)
- Make a 20 mg/ml (100 X) stock solution of INK128 in NMP. Freeze aliquots and store at -80.
- Prepare vehicle solution---15% PVP/H2O: Dissolve 15.8 g of PVP into 84.2 mL of water and then sterile filtered. Once this solution is made, keep in dark at room temperature.
- Make final INK128 solution: Dilute stock 1:100 into 15% PVP/H2O for final concentration of 0.2 mg/ml. Dosing solutions seem to be fine for 2-3 weeks at 4 C kept in the dark.
- Dose at 100 ul po/20 g mouse (=1 mg/kg).
Note: Only after dissolving drug in NMP should then combine with the 99% PVP/ H2O solution.
- INK128 is from LC Laboratories, product number I-3344, 10mg. Link: http://www.lclabs.com/products/i-3344-ink128-free-base
- NMP is from Sigma-Aldrich, product number #328634, 100ml (1-Methyl 2-pyrrolidinone, anhydrous, 99.5%). Link: http://www.sigmaaldrich.com/catalog/product/sial/328634?lang=en®io n=US
- The PVP is from Sigma-Aldrich, product number #81420, 100g (Polyvinylpyrrolidone K 30 avg MW 40,000). Link: http://www.sigmaaldrich.com/catalog/product/sial/81420?lang=en®ion =US
For in vitro experiments
- Purchase INK128 with the amount of 1mg.
- Dissolve INK128 in 3.23 ml of DMSO to the stock concentration of 1mM/L. For long term storage, freeze aliquots and store at -80.
- Then dilute INK128 to a middle concentration with cell culture medium before adding to cell culture.