Hello!
I am planning to use Bradford assay to measure protein content in PBMCs lysates. The problem is that Bradford assay doesn't seem to be compatible with Tris-based buffers or SDS. Are there any other methods to extract total protein from PBMC suspension that doesn't include these compounds or high temperature?
Thanks!
You could try the Pierce Coomassie Protein assay, it's a modified version that tolerates tris (2M) and you could substitute CHAPS for the SDS as the assay tolerates 5% vs 0.125%,. You should be able to compare efficiency empirically by running a gel with your SDS extraction vs CHAPS extraction.
Thank you, Stanley Ng, I definitely should try to switch to Pierce assay if this one won't work out. The idea with CHAPS sounds good as well, thanks a lot!
Hello Violetta
Try diluting your samples in water say at 1:5 or 1:10 dilution and use the diluted samples for Bradford assay. Give it a try may be it will work.
Regards.
Hope the following links will be helpful:
https://www.researchgate.net/post/Which-lysis-buffer-works-best-with-Bradford-reagent
https://www.researchgate.net/post/Any_good_method_to_extract_total_protein_from_PBMC_for_my_western_blot_analysis
Thank you, Malcolm Nobre
I've recently tried to dilute my sample in order to decrease the impact of lysis buffer. Unfortunately, it didn't help much so I guess I'll have to switch to another method.
Thanks for your advice!
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