Login

Join for Free!
25818 members


bradford assay of plant protein

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

Moderator: BioTeam

bradford assay of plant protein

Postby aravn22 » Wed Jan 28, 2009 9:40 am

this may be really simple but i have no experience in dealing with proteins.
I have a crude extract (plant protein) and i want to do bradford assay but i don't know how to proceed with it. The protocol i have says that i dilute my standard (bovine gamma globulin, BGG) and the crude protein extract in 0.15 NaCl. other protocols i read, however, use water or buffer. So which should I use? I extracted the plant protein using pvpp, mops, dtt, edta, mgcl2 and pmsf. do i have to use the same buffer to dilute standard and sample for bradford?
aravn22
Garter
Garter
 
Posts: 1
Joined: Wed Jan 28, 2009 9:29 am

Postby GreenDog » Thu Jan 29, 2009 1:19 pm

You can use water. 0.8mlDDw+0.2mlBSA+4microliter of desired protein. If you are over the scope of your standard curve or spectrometer dilute your protein 1/5 or 1/10, or whatever you like.
Make standard curve with the BGG 1,2,4,8, microgram (or something similar).
Follow the instructions i.e. don't forget the blank, make all your samples at the same time, mix well, and wait for 15 min before reading.
"When In Danger Or In Doubt Run In Circles Scream And Shout"
Lawrence J. Peter
User avatar
GreenDog
Coral
Coral
 
Posts: 126
Joined: Sun Mar 27, 2005 8:37 am


Return to Molecular Biology

Who is online

Users browsing this forum: No registered users and 0 guests