detection of proteins in FPLC

Tom Chappell t.chappell at ucl.ac.uk
Thu Mar 20 11:10:22 EST 1997


In article <E7BFFy.Bpz.B.midge at bath.ac.uk>, bspwrb at bath.ac.uk (W R
Bennett) wrote:

>In the referenced article, dalmiabk at phibred.com (bipin k. dalmia) writes:
>>On Tue, 18 Mar 1997 21:49:38 GMT, bspwrb at bath.ac.uk (W R Bennett)
>>wrote:
>>
>>>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. 
>>
>>this would assume a flow-cell path length of 1.0 cm and i do not think
>>that detector flow-cells have a 1.0 cm path length.
>>
>Good point, but I think the flow-cell that came with our Pharmacia UV-M
>(ten years old, admittedly) did actually have a 1.0cm path length, but
>it's easy to check - just look in that huge binder you get with the FPLC
>system to see what your path length is. 
>
>Bill Bennett
>Developmental Biology Unit,
>School of Biology and Biochemistry
>University of Bath
>
>bspwrb at bath.ac.uk

Correct me if I'm wrong on this, but isn't an AU defined as absorbance /
distance and thus independent of path length? If you are getting
absorbance as an output, you have to take path length into consideration,
but if you are getting AU's, the electronics or software should already be
taking the path length into account.




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