Suppose you had 85 of a protein sample. Of this 12 1 were diluted into 108 1 of

Question

Suppose you had 85 of a protein sample. Of this 12 1 were diluted into 108 1 of water. Of this dilution 251 were submitted to Bradford protein assay and produced art OD 0.63) This is a thought question: for pure molecules, one would Slat 0 1220 so use the Lambert-Beer equation and need the extinction coefficient. But, somewe biochemical assays do not contain pure molecules under defined conditions, thereby require a standard curve. If you have been doing this lab, you already have a prio standard curve, where? yo0 32l ,7. a. What is the dilution from the originaf sampte that 25 ul measured in Bradford assay represents? (2 points) b. How mu rotein is in the original protein sample? (3 points) 6 3 Dw how to do Please leave 14-for Some,na e iSe

Explanation / Answer

Solution:

The orginal protein sample was diluted 1/10 times. This is done by mixing 12 ul of original protein sample with 108 ul of water. The dilited sample gave an OD of 0.63.

The microplate Bradford assay achieves an absorbance of 0.1 at about 6 micrograms/ml

So the protein concentration is 0.63* 10 * 6 micrograms/ml

= 37.8 micrograms/ml is the concentration of the protein or

= 0.037 micrograms/ul

So for 85 ul it would be 0.037 * 85 = 3.145 micrograms

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