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protein quantitation


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3 replies to this topic

#1 Nay

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Posted 30 March 2005 - 06:18 PM

I am after a protein quantitation method which is ok to use when there is detergents in your solubilisation/lysis buffer. I would normally use the Biorad protein assay (Lowry assay), but I am getting massive errors (always increased estimates) of my protein concentrations. I have been using trizol to extract the protein so I assume it is due to a carry-over of phenol (when I don't use trizol, my lowry assay works well) - is there a quantitation assay which I can use if I have some phenol contamination?

I am sick of trial and error loading! :)

#2 ros

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Posted 31 March 2005 - 06:01 PM

I am after a protein quantitation method which is ok to use when there is detergents in your solubilisation/lysis buffer.  I would normally use the Biorad protein assay (Lowry assay), but I am getting massive errors (always increased estimates) of my protein concentrations.  I have been using trizol to extract the protein so I assume it is due to a carry-over of phenol (when I don't use trizol, my lowry assay works well) - is there a quantitation assay which I can use if I have some phenol contamination?

I am sick of trial and error loading!  :(

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this might not help - but do you have to use Trizol? in my experience, Trizol is more trouble than it is worth for both RNA and protein! I would suggest changing your isolation protocol.

#3 Nay

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Posted 31 March 2005 - 06:21 PM

my antibody is horribly fussy and only seems to work when I use trizol to extract my proteins - I have decided to try dialysing my samples and/or using a buffer exchange column before I quantitate them now...

#4 neuron

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Posted 30 January 2009 - 07:50 AM

you can try this-


http://biology200.gs..... protocol.pdf


Hope it helps ;)




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