NaH2PO4 20- 50 mM

NaCl 500- 1000 mM

Sorbitol 200 mM

PI(Sigma) 1:100

PMSF 1:100

DNAse(1mg/mL) 1:100

MgCl2 (1M) 1:100

Triton X114? (5%-10%?)

Depending on the stability of proteins Triton-X114 might be helpful

If you add proper amount of Triton-X114 to your extraction buffer, it will separate above 22°C into two phases so that you will have in dense phase mainly membrane proteins which are not soluble in water while the less dense phase will include less detergent and mostly water-soluble proteins

Transmembrane protein can be extracted with sodium deoxycholate. Cytochrome c oxidase is an example, that could be extracted sodium deoxycholate

Would a milder detergent such as dodecyl maltoside work?

Thank you! Bharatratnam Parayitam Do you have a protocol for your method?