A student has been given a sample of human blood and instructed to measure the c

Question

A student has been given a sample of human blood and instructed to measure the concentration of hemoglobin in it. He realizes that the absorption is so strong that he will have to dilute it in order to obtain a reliable measurement. After a few tries, he discovers that a sample diluted 1000-fold gives an absorbance of 1.4 at 420 nm, in a cuvette with a path length of 1 cm

Another students suggests that it might be a good idea to measure the hemoglobin concentration using the Bradford dye-binding assay.

a. What advantage might the Bradford assay have for this situation?

b.What disadvantage might the Bradford assay have for this situation?

Explanation / Answer

a) It is done in one step where the Bradford reagent is added to a test tube along with the sample. After mixing well, the mixture almost immediately changes to a blue color and the absorbance can be read it.

The Bradford protein assay can measure protein quantities as little as 1 to 20 g It is an extremely sensitive technique.

b). in dilute sample the Bradford assay is linear over a short range, typically from 0 µg/mL to 2000 µg/mL, In making these dilutions, error in one dilution is compounded in further dilutions resulting in a linear relationship that may not always be accurate.

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