Protein Measuring: Standard vs Sample Solutions

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In the discussion, the preparation of standard and sample solutions for protein concentration measurement is outlined. Standard solutions consist of 800 µL water, 20 µL of each standard solution, and 200 µL of dye concentrate, while sample solutions include 800 µL water, 4 µL of each sample solution, and 200 µL of dye concentrate. A key question arises regarding the interpretation of a measured concentration of 0.5 mg/mL for a sample. It is clarified that this value reflects the concentration based on the 4 µL sample volume, and adjustments should be made to express the concentration consistently, typically per mL. The discussion emphasizes that the standards serve as a reference for quantifying the samples, and all calculations should align to a common scale to ensure accuracy in reporting protein concentrations.
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When I make the standard curve then each standard solution is prepared like this:
800ul water
20ul each standard solution
200ul dye concentrate


The samples are prepared like this:
800ul water
4ul each sample solution
200ul dye concentrate

I wonder if I for instance get 0.5mg/ml on one of my sample. Does this mean that this value is the concentration of the original sample or should I multiply with 5 since the volume of the standard is larger (20ul) then the volume of the sample (4ul)?
 
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Could you be more specific of how you are measuring your proteins?
Are you using a spec?
Usually the standards just standardize your samples, a ladder to measure against.
The only thing you should be compensating for is the volume you put in. You are measuring the concentration /4ul. This then should be calculated so you have a value either per ul or per ml. Its hard for me to say exactly because I'm not sure how you are measuring the protein concentration. Usually the standards tell you the amount of protein you are adding and everything should be calculated to be on the same scale (eg how many ug/ml and not ug/5ml for one and ug/3ml for another sample).
 
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