Ruchi can you please tell whether you are trying to obtain from the cell line or tissue??

Fatemah, there is a different protocol to lyse for tissues, because in case of tissues you need to crush the tissue properly otherwise it wont get lysed !!

and ya Fatemeh, dont forget to use protease inhibitors, Ruchi, do u use protease inhibitors in case of your lsis buffer?

Hi Dipon,

I m trying to obtain the protein from an endometrial cell line. I do not use any protease inhibitor but i had tried adding PMSF while preparing a lysate but could not get the protein of interest

Ruchi, I worked with Rab proteins and used RIPA buffer (without SDS). that's works very well.