At least with large transmembrane proteins it turns out that results are often better if samples are not boiled, but heated to only 60 °C. I am not aware of any systematic studies on this, it is simply an experience that many have made, including myself. Gentle does it!
Engelbert Buxbaum : Dr i have another question that i require your advice if possible https://www.researchgate.net/post/i_cant_get_protein_signals_using_RIPA_buffer_for_extraction_of_one_brain_hemisphere_myhous_keeping_ptn_is_cytoplasmic_ptn_beta_actin_no_signal?isAnswerFieldFocused=true
Heating is not necessary if proper sds conc is used, perhaps 15 min at 37 C works for all proteins even membrane ones. The hydrophobic effect increases with temperature, this is why you should not boil membrane proteins.
- Badges
- Science method