Trouble with Bio rad protein assey

4 posts / 0 new
Last post
Dtvt2001
Dtvt2001's picture
Trouble with Bio rad protein assey

I am doing a ChIP assay, I am checking my samples protein concentration with the bio rad protein assay. When i place my sample into my bio rad solution, the protein seems to clump together. I treated my cells with either DMSO or TCDD for 1.5H. Then I add 1% formaldehyde for ten mins. Then I washed with one Ice cold pbs. Next i peleted my cells and remove the extra pbs and store in -80. Then i attempted to do the bio rad protein assay and Thats where i got my problem.

on a side note my cells were grown in 100mm plates and at 100 confluency.

Any help would be greatful

Dtvt2001
Dtvt2001's picture
also my lysis buffer is 1%sds

also my lysis buffer is 1%sds,10mM EDTA,50mM Tris-HCL. 2ul of sample in to 998ul of biorad solution

bwbrian
bwbrian's picture
If I remember right, you can

If I remember right, you can't use the BioRad assay with detergent, and you mentioned using 1% SDS in your lysis buffer. You might try the BCA Assay instead because it works with detergents.

Eric Larson
Eric Larson's picture
Bio-Rad has a DC Protein

Bio-Rad has a DC Protein Assay which is detergent compatible (DC).  It can handle up to 10% SDS.  Info on the web site, www.bio-rad.com.