Does trypsin-based cell harvesting damage membrane proteins?

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clavin23
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Does trypsin-based cell harvesting damage membrane proteins?

Post by clavin23 » Tue Mar 20, 2007 11:01 am

I am trying to get cell lysates to study the membrane-bound receptor EGFR. I am using trypsin to harvest adherent cells from a 15 cm flask, spin the cells, remove the trypsin buffer and then I lyse the cells with a an NP-40 plus protease inhbitors buffer. My question is does the trypsin damage (cleave) my EGFR protein? If I use 8 M urea to lyse the cells is it any different?
Thank you in advance.

Vagabond
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Post by Vagabond » Tue Mar 20, 2007 12:48 pm

Yes Trypsin will damage a lot of membrane bound proteins. I don not know if EGFR itself has a Trypsin recog site though. You could try to physically scrape the cells off.
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