bradford assay of plant protein

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bradford assay of plant protein

Post by 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?

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Post by 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"
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