Solve Braford Assay: Why is My Answer 20X Different?

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The discussion centers on a misunderstanding of the Braford Assay calculations, where the user initially calculated a protein concentration of 0.00842 µg/µl, while the correct answer is 0.168 mg/ml, differing by a factor of 20. The discrepancy arises from the dilution factor, as the sample was diluted 20 times to a total volume of 1000 µl. The user realizes that to find the protein concentration, they must account for the dilution, leading to the correct concentration calculation of 0.1684 mg/ml. Clarifications about unit conversions and dilution factors are crucial for accurate results in such assays. Understanding these concepts resolves the confusion regarding the factor of 20 difference in the answers.
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Homework Statement



https://www.physicsforums.com/attachments/76905
https://www.physicsforums.com/attachments/76906

Homework Equations

3. The Attempt at a Solution [/B]
Hi I have a question on Braford Assay:
I am given this question:
I did the caluclation as follows:
y = 66.717x
Since given absorbance = 0.562
0.562 = 66.717x
x = 0.00842ug/ul
But the answers given is 0.168mg/ml
I do not know what I did wrongly? Can anyone please help? Thanks!
My answer and the correct answer differ by a factor of 20? Why is that so?
 
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tvtokyo said:
My answer and the correct answer differ by a factor of 20?

0.562 is not absorbance on the original solution.
 
Yes, I was trying to answer this question but when I press your links this time I am told I don't have right to access the page. But I remember when I saw it before there was something about an 0.05 ml sample - which is 1/20 of an ml. Could these occurrences of 1/20 be in any easy connected?
 
I still don't get it though. Sorry. I will post the image again, Sorry for the trouble pal!
Anyway, even it is a factor of 20. My answer has a unit of ug/ul and the correct answer is in mg/ml so technically it is not a factor of 20??

upload_2014-12-26_10-21-24.png

upload_2014-12-26_10-21-47.png
 
Please reread the last paragraph of the attached text, it clearly tells you why the difference is 20.

50+750+200=1000.
 
I am suppose to find the protein concentration of the liver sample what do i do with the 1000ul ?
Is this correct?
Since x = 0.00842ug/ul and total volume of my assay --> 1000 ul
Then mass = 0.00842 * 1000 = 8.42 ug
Concentration of protein = 8.42ug/50ul (of protein in liver)
= 0.1684 ug/ul = 0.1684 mg/ml
 
Correct.

50 μl of your sample were diluted 20 times to 1000μl.
 
Thank you!
 
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