berrycom, on Nov 17 2009, 05:15 AM, said:
TracyDuke, on Oct 19 2009, 01:54 PM, said:
. Click on the first box that says" cell separation" on the upper left corner on the mainpage. They have positive and negative selection reagents and kits, depending on your needs. A lot of literatures use their products.
Many thanks for your reply. Actually, I have tried Miltenyi lineage depletion kit but I had a viability issue. After one day of culture approximately 60% cells were dead (trypan blue exclusion). Has anybody used this kit and cultured the hematopoietic progenitors afterwards? I had many samples so the selection process was quite lengthy. Maybe that was the problem. I need to repeat it.
I have some experience with miltenyi lineage depletion kit, to be specific: CD133 microbead isolation kit. I also have some issue with the viability and purity. To improve the viability, I have increase the BSA to 5-10 % in the buffer plus make sure that the buffer is cold during the isolation procedure been carried out. And to obtain the purer population, I run down the cells suspension thru the isolation column twice. So far it is work.
But recently I talked to one competitor who is selling the isolation product also. he said isolation kit from miltenyi using dextran particles/beads to bind to cells that might interfere when you are about to identify/characterize the isolated cells using flow cytometry. whereas theirs, using nano particles/beads that wont interfere in fact, flow cytometry couldnt able to identify/ pick up the particles. i do not know how true it is..i havent really find out about it yet..