Calculating Protein Concentration w/ Bradford Assay

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SUMMARY

The discussion centers on calculating protein concentration using the Bradford Assay, specifically with a 5 times dilution of protein A. The standard curve equation derived is y = 66.717x, leading to an absorbance reading of 0.497. The initial calculation yields 0.0372 mg/ml, but the correct concentration is identified as 0.372 mg/ml. The discrepancy arises from a misunderstanding of the dilution factor and its application in the final concentration calculation.

PREREQUISITES
  • Understanding of the Bradford Assay methodology
  • Familiarity with standard curve calculations in spectrophotometry
  • Knowledge of dilution factors and their impact on concentration
  • Basic mathematical skills for unit conversion (ug/ul to mg/ml)
NEXT STEPS
  • Review the principles of the Bradford Assay and its applications in protein quantification
  • Learn how to construct and interpret standard curves in spectrophotometric assays
  • Study the effects of dilution on concentration calculations in laboratory settings
  • Explore common pitfalls in protein concentration calculations and how to avoid them
USEFUL FOR

Students in biochemistry or molecular biology, laboratory technicians performing protein assays, and researchers analyzing protein concentrations using the Bradford Assay.

tvtokyo
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Homework Statement


Hi, I have one more question about the Bradford Assay. This one involves dilution.

Given 1ml of protein A, I am to determine its protein concentration (in mg/ml) using Bradford Assay (with known BSA protein standards).
Also, Under a 5 times dilution, the average absorbance is 0.497.

Homework Equations


The standard curve I obtained is:
upload_2014-12-26_10-35-0.png


The Attempt at a Solution


From the standard curve,
y = 66.717x
With absorbance = 0.497
0.497 = 66.717x
x = 0.00744 ug/ul
Since there is a dilution factor of 5
New x = 0.00744 * 5 = 0.0372 ug/ul = 0.0372 mg/ml ?? (is this correct?)

However, the correct answer is 0.372 mg/ml . What did I did wrongly??
 
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tvtokyo said:
x = 0.00744 ug/ul
Looks good.
tvtokyo said:
New x = 0.00744 * 5 = 0.0372 ug/ul = 0.0372 mg/ml ??
Looks good.

Answer keys in textbooks are "edited/proofed" by "slave labor," aka grad students and post-docs. Often, "half" changes are made after proofing --- problem statements change, and answers don't.
 
In your other question you clearly ignored part of the information given, hard to comment on this one not seeing the whole problem.
 
Is it another part of the same problem? Might be best in future to get all the questions together before asking for help.
 

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