Bradford protein determination; calibration curve

H.V.Taylor harold.taylor.remove-this at
Sat Dec 18 11:26:46 EST 1999

PGegen at (Dr. Peter Gegenheimer) wrote:

>> You'll find that fitting your results to a hyperbolic function 
>> i.e.,  Ext. = (protein * factor1) / (protein + factor2)
>> works quite well.
>If the relationship is not linear, you would first suspect that the reaction 
>is not going to completion; that is, reagent concentration is limiting. This 
>is especially true for the Bradford assay, which is (in principle) a simple 
>binding isotherm. 
>[On the other hand, a curve of Abs vs [dye] must have a hyperbolic shape 
>because its a binding isotherm. From this you could measure Kd but you 
>wouldn't want to use it for a calibrations curve of dye concentration!]
Gee, I wish I'd put it that way ;-))

Thanks for cleaning up after me. Merry Christmas all from Germany!


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