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Bradford Protein Assay question - Protein Science

Bradford Protein Assay question - Post Questions and Discuss Protein Structure and Function, Protein Localization etc.. Please do not post Proteomic questions here, there is a Proteomic Forum below.

#1
12-17-2007, 07:25 PM
Pipette Filler

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Ok, this seems like a simple question, but I am getting different answers from different people. when I do a Bradford protein assay, I dilute my samples 1:10. When I get my adjusted average, should I multiply that by 10 BEFORE putting it in the equation from the standard curve or do I plug in the adjusted average and then multiply that answer by 10? The answers are not the same according to my calculator. And is there a place that I can find the answer I get so that I can use it as a reference when someone asks? Thanks for reading!
#2
12-17-2007, 09:44 PM
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Protein samples are diluted 1:10 not the standards, therefore you use the standards to calculate the concentration ug/ml or ug/ul of the diluted samples, then you correct for dilution factor by multiplying by ten. The point is not to multiply the Absorbance values, or the standards.
#3
12-19-2007, 02:11 PM
Pipette Filler

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Quote:
 Originally Posted by danfive Protein samples are diluted 1:10 not the standards, therefore you use the standards to calculate the concentration ug/ml or ug/ul of the diluted samples, then you correct for dilution factor by multiplying by ten. The point is not to multiply the Absorbance values, or the standards.
I understand that I do not dilute the standards. I want to know if I multiply the absorbances of the samples by 10 before I put them into the equation I get from the standard curve?
#4
12-19-2007, 02:49 PM
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Quote:
 Originally Posted by klhayes I understand that I do not dilute the standards. I want to know if I multiply the absorbances of the samples by 10 before I put them into the equation I get from the standard curve?
No, Do not multiply the Absorbance values, use the avg abs to calculate the protein conc of the dilution then multiply times the dilution factor.
#5
11-17-2008, 03:57 AM
Pipette Filler

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I agree with Danfive.

BTW, I am either blind or there is no post new thread in here. I will then put my request here, I am hoping to get info on an easy way to remove albumin from a mAb with a total volume of 100 ul. I appreciate any comment.
#6
01-06-2009, 09:58 PM
Pipette Filler

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Pls, i Just join forum, are you talking about the dilution factor for the preparation for the protein assay and in what analysis and to what respect. pls be specific. pls comment

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