Login

Join for Free!
119208 members


plasma protein precipitation

Discussion of all aspects of biological molecules, biochemical processes and laboratory procedures in the field.

Moderator: BioTeam

plasma protein precipitation

Postby varshaparekh » Sat Jul 12, 2008 3:35 pm

Hi all,
Does anybody have a protocol for precipitation of plasma proteins from human blood
varshaparekh
Garter
Garter
 
Posts: 2
Joined: Sat Jul 12, 2008 3:29 pm

Postby victor » Wed Jul 16, 2008 2:02 am

1. Use RBC lysis Buffer about 3x blood volume and incubate for 10 minutes and centrifuge at 1500 rpm; 10 minutes (rt) and after that, pour the supernatant --> do this 3 times.
2. Vortex the pellet and use Cell lysis solution (about 625 uL for 2,5 mL blood) & 2,5 uL RNase --> incubate for 30 minutes; 37C.
3. Do the protein precipitation by adding 5M ammonium acetate (about 417 uL for 2,5 mL blood sample) and vortex it until the solution looks white milky.
4. Centrifuge at 3000 rpm, 4C, 15 minutes and pour the supernatant.
5. There you'll get blood protein and plasma protein.

Hope this helps.
Victor
Q: Why are chemists great for solving problems?
A: They have all the solutions.
User avatar
victor
King Cobra
King Cobra
 
Posts: 2668
Joined: Sat Apr 30, 2005 12:01 pm
Location: Yogyakarta, Indonesia..


Return to Molecular Biology

Who is online

Users browsing this forum: No registered users and 1 guest