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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.

Sincerely,

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closed as unclear what you're asking by canadianer, David, kmm, Bryan Krause, WYSIWYG♦ Jan 9 at 10:50

Please clarify your specific problem or add additional details to highlight exactly what you need. As it's currently written, it’s hard to tell exactly what you're asking. See the How to Ask page for help clarifying this question. If this question can be reworded to fit the rules in the help center, please edit the question.

  • $\begingroup$ As @Price mentioned, it doesn't make sense to talk about number of cells in a lysate. Lysis breaks the cell membranes, so there are no cells after lysis. Are you trying to determine how much volume of lysate represents the contents of N cells? $\endgroup$ – divibisan Dec 21 '18 at 23:54