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.
Hi Basma,
With your describe proteins you should go straight forward to 95C for 10 minutes atleast.
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