detection of proteins in FPLC

W R Bennett bspwrb at bath.ac.uk
Tue Mar 18 16:49:38 EST 1997


In the referenced article, ita at DNA.CBIOT.UFRGS.BR (ITABAJARA DA SILVA VAZ JUNIOR) writes:
>Please,
>Is there a simple method of transform the AU detected by  monitor UV in FPLC 
>in mg/ml concentration?
>Thanks, 
>
Assuming 280nm detection, 1.00 AU is roughly equivalent to 1.0 mg/ml, so,
for example, if your sensitivity is set to 0.01, then full scale
deflection is roughly 10ug/ml.  The total amount of protein in a pooled
peak is proportional to the area under the peak, but if you don't have an
integrating chart-recorder, as a rule of thumb, we used to assume that,
when we pooled a bunch of fractions from an FPLC run, the protein
concentration of the pool was about half the height of the peak, (since
most of the fractions in the pool are not the full peak height).  
Obviously, this starts to break down if you start adding in loads of 
low-protein fractions from trailing edges of the peak.

Cheers,

Bill Bennett
Developmental Biology Unit,
School of Biology and Biochemistry
University of Bath

bspwrb at bath.ac.uk



More information about the Methods mailing list