I am working on drug targeting studies. I want to check the amount of drug uptake. From total cells first I want to separate and collect the cells which have taken up the drug. The method reported has written the use of 70% of Percoll. I want to know why 70% of density gradient separator is used? How has it been decided? What is the basic mechanism behind it?
Percoll gradient separator works on the principle of Isopycnic separation
In this type of separation, a particle of a particular density will sink during centrifugation until a position is reached where the density of the surrounding solution is exactly the same as the density of the particle. Once this quasi-equilibrium is reached, the length of centrifugation does not have any influence on the migration of the particle.
http://www.coleparmer.com/TechLibraryArticle/30
Percoll gradient separator works on the principle of Isopycnic separation
In this type of separation, a particle of a particular density will sink during centrifugation until a position is reached where the density of the surrounding solution is exactly the same as the density of the particle. Once this quasi-equilibrium is reached, the length of centrifugation does not have any influence on the migration of the particle.
http://www.coleparmer.com/TechLibraryArticle/30
The answer for Percoll can be found by referencing the work of Boyum who pioneered the technique. You might also look into separation of cells using antibodes or Dynabeads which separate cells by magnetic atraction.
dr Sonali, which centrifuge machine are you going to use for this method of percoll density gradient ? a simple one or an ultracentrifuge? can anybody please answer?
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