I am planning to isolate Islets from a rat pancreas using Ficoll density gradient of 1.108g/mL,; 1.096g/ML; 1.069g/mLand 1.037g/mL. I have bought Ficoll 400 powder and I would like to make these solutions. Is there anybody out there who has the protocol showing cuculations, formulas of how to make these solutions? 1.108g of Ficoll cannot dissolve in 1 mL of deionised water or buffer even when you heat it.!!!! The data sheet indicate that solubility of Ficoll is 1 g in 10 mL. So, how can one make a solution of 1.108g/mL?
According to the GE Healthcare web page about Ficoll400, it is soluble up to 50% (i.e. 50g/100ml of solution), resulting in a density of 1.2 g/ml. This is the density of the solution, not the concentration of the Ficoll. Remember, water has a density, too. (1g/ml at 4oC for pure water). If you can't find a table for how to make solutions of various densities, you could make this 1.2g/ml solution, then dilute it with water (or buffer) until it has the desired density, which you could determine by weighing 1 ml on an analytical balance.
Thanks Adam. I will try. nIf I stillhave problems I will come back to you
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