In general, most protein quantification methods
have significant problems with idiosyncracy, because proteins are
quite variable in composition and structure. The best method is to
use is the absorption at 280 nm, but this is quantitatively useful
only if the
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--- On Sat, 9/4/11, Arpit Mishra ar...@igib.in wrote:
From: Arpit Mishra ar...@igib.in
Subject: [ccp4bb] how to quantitate protein which dont have ne aromatic
Hi,
You can read the spectrophotometric absorption at 280 nm and 200nm in UV
range.
It should serve your purpose and provide a decent idea for the amount of
protein in the sample.
Provided that absorbance at 280nm is given by aromatic rings but at the same
time absorbance at 200nm is contributed
hello everybody
i am working on the protien which dont have any aromatic residue i do fplc
other purification using 220 absorption, but i want to quantitate protein
precisely i have tried using BCA nd bradford but both methods quantification
is not matching,,so any one is having sum idea how to
you can do amino acid analysis on your pure protein, using a commercial or
academic service - I hope these are still around. You should only need to do
this once, then relate the result to your A220, BCA and Bradford assays.
Mark
Quoting Arpit Mishra:
hello everybody
i am working on the
: [ccp4bb] how to quantitate protein which dont have ne aromatic residue
hello everybody
i am working on the protien which dont have any aromatic residue i do fplc
other purification using 220 absorption, but i want to quantitate protein
precisely i have tried using BCA nd bradford but both
:00 US/Canada Pacific
Subject: [ccp4bb] how to quantitate protein which dont have ne
aromatic residue
hello everybody
i am working on the protien which dont have any aromatic residue i
do fplc other purification using 220 absorption, but i want to
quantitate protein precisely i have
At 9:47 AM -0700 4/9/11, Michael Thompson wrote:
Bradford dye binds to hydrophobic residues, mainly aromatics,
The statement above is not accurate.
Compton and Jones. Anal. Biochem. 151(2): 369-374, 1985
- John
--
It is not surprising that your bradford and BCA assays don't agree if you
have no aromatic amino acids in your protein. Bradford dye binds to
hydrophobic residues, mainly aromatics, so I would guess your bradford is
consistantly giving lower measurements than the BCA assay. I also wouldn't
be
.
Georgiou, et al. Anal Bioanal Chem. 2008 May;391(1):391-403.
Cheers,
Mike
- Original Message -
From: John A. Newitt newit...@gmail.com
To: CCP4BB@JISCMAIL.AC.UK
Sent: Saturday, April 9, 2011 12:06:23 PM GMT -08:00 US/Canada Pacific
Subject: Re: [ccp4bb] how to quantitate protein which
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