I intend to do CD34 cell sorting so how much cells do I require? Also what is the required amount of proteins fiom these same cells to do disease protein profiling using protein array?
Thanks in advance.
Wei Xing
Dear Xing,
To be precise-it varies from cell to cell type and yield of cells !
Minimum number of cells required for a sorting depends on percentage of cells in your sample and how many cells you want to isolate. e.g. for side population analysis generally 1-2% population is SP, so you may need to go for approximately 4-6 T-75 flask's trypsinization to get sufficient number of cells for applications like RNA isolation. so if you get a decent 40-50% cells from your sample then sorting from only 1-T75 flask with approximately 4-5 million cells should be sufficient to give you decent protein for protein array analysis.
Hope it helps !
What type of samples are we talking about: Blood, bone marrow or something else? Usually/unfortunately the only way to find out is to simply do one test sort ;-)
Thanks Ajay, it certainly is of help.
Anouk, the samples are actually differentiating myeloid cells generated from leukemia iPSC. My intent is to separate progenitors from mature forms using flow-cytometry, followed by protein array analysis. I'm thinking of performing a differential protein expression between my samples with patient bone marrow as a control as well. How does that sound?
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