IUBio

Bradford protein determination; calibration curve

H.V.Taylor harold.taylor.delete-this at remove-this.uni-tuebingen.de
Fri Dec 17 11:57:13 EST 1999


>P. Oranje wrote in message
>> At the moment I am purifing peroxidase from horseradish
>> roots. Protein determination is performed according to
>> Bradford's assay. Can someone tell me what the general
>> mathematical formula is that describes the correlation
>> between protein concentration and extinction best when
>> using Bradford's assay for protein determination ?
>
>What are you using to calibrate your protein concentrations?  Why not use
>Horseradish peroxidase, that you have bought from Sigma! 

You'll find that fitting your results to a hyperbolic function 

i.e.,  Ext. = (protein * factor1) / (protein + factor2)

works quite well.

Harold





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