The goal of these types of experiments is global quantification of proteins expression. This can be done either label free or using isotopic labeling. We use the S-trap method for all lysates.
Regardless of the quantification method, our preference is:
- Cell lines: receive a cell pellet. For ‘medium coverage’ we need minimum of 1e5 cells. For ‘deep coverage’ 5e5 cells. We do have workflows for much lower cell numbers - please consult with us about this.
- Tissue: protein extraction must include physical grinding using bead-beating, homogenizer or mortar and pestle in the lysis buffer. Our current preference is 5%SDS, in Tris ph 7.4. We require 20ug protein for ‘medium coverge’, 200ug for ‘deep coverage’.