A couple questions about the bradford method

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SUMMARY

The Bradford method requires two blanks of 0.15M NaCl to ensure accurate photometric measurements. One blank is used to zero the photometer with water, while the second blank is prepared with Coomassie Brilliant Blue to measure absorption. This approach allows for the subtraction of the blank's absorption from the standards when plotting the standard curve. Additionally, serial dilutions of the unknown sample are necessary to ensure that the concentration falls within the detectable range of the assay.

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Homework Statement



In the protocol for the bradford method I am told to prepare two blanks of water of .15M NaCl. I don't understand why I need two of them. Don't I just need one of them for placing into the photometer after I have set 0% transmission to set 100% transmission by.

should I put some of the coomassie brilliant blue, in one of these, both of them, or none of them?

I really don't get why I need two of them, any help would be appreciated





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The Attempt at a Solution

 
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Is it that one of the blanks is left with water to zero the machine and the other blank has just coomassie brilliant blue added to it, measured for absorption, and then subtract the absorption from the standards when plotting the standard curve, because that is what I am told to do for my Bicinchoninic acid assay.

Also, why do I have serial dilutions of the unknown, is it just in case the stock solution of unknown falls outside of the range of what concentrations are detectable by my test.

Furthermore, if I am running a Bicinchoninic acid assay do I have to do serial dilutions just as I did for the bradford method.
 

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