I would like to know that how can we take low or precise number of cells from a cell lysate? (without using any protein quantification assay).
Let say, If I have a cell line having cell density 5x10^5 cells/mL (and its avg amount of protein concentration is 0.1ng/cell)
If I lyse these cells at the concentration of 5x10^5 cell/mL with buffer (eg., 0.1% rapigest + 10mM DTT). After achieving cell lysate, how can I take precise cell numbers from this lysate for further sample processing (e.g, alkylation, digestion etc)?
For eg., if I want to have 100cells/5ul or 1000cells/50ul , then how can I take it from cell lysate?
I know I'll have to dilute my lysate, but I'm bit confused how to achieve that based on cell concentration? how much lysis buffer and dilution would be appropriate?
Note: In my case I can't use any protein quantification assay for measuring protein concentration.
I would highly appreciate if anyone can help please.
Looking forward to hear soon.