Chemical Forums

Specialty Chemistry Forums => Biochemistry and Chemical Biology Forum => Topic started by: joedassin on June 10, 2012, 12:17:28 AM

Title: Protein estimation by the Bradford assay
Post by: joedassin on June 10, 2012, 12:17:28 AM

Hello everyone, I have a question concerning intro to biochem class.  I am suppose to plot the standard curve (absorbance on y axis and concentration of protein on the x axis). I used BSA (2.0mg/ml) to make 7 different BSA standards. For example, for the first standard, I have 2.0 ml of Bradford reagent, 50 microlit of distilled water and 20 microlit of protein sample. I have my absorbances values but I dont see where I have to take the concentration from (to plot on the graph)?

Thank you kindly for your help
Joe

Title: Re: Protein estimation by the Bradford assay
Post by: Arkcon on June 10, 2012, 06:23:46 AM

Hmmm ... so you can plot your known concentrations vs observed absorbances, right?  You gave an example, for your first standard, so you can just do that for the rest.  Maybe you have another question?

Title: Re: Protein estimation by the Bradford assay
Post by: Babcock_Hall on June 11, 2012, 09:30:30 AM

Are you having trouble with the calculation of protein concentration in the cuvette?  This is a dilution calculation, but my experience is that students often stumble over dilution calculations.