I n Proteomics Practicals a) Why is it necessary to determine the amount and con

Question

In Proteomics Practicals

a) Why is it necessary to determine the amount and concentration of protein in a sample extract? (short answer)

b) If you use a number of different assays to determine the amount of protein in a sample, would you expect the value to be exactly the same? Explain your answer.

c) If you use a number of different assays to determine the amount of protein in a sample, would you expect the relative values between samples to be the same? In other words, if two samples had a relative total protein amount of 1:2 (twice as much in sample 2) in one assay, would the relative total amount determined by another assay be 1:2?

Explanation / Answer

a. Protein is the most important biochemical molecule which form the body mass. These proteins are present in many forms like peptide, polypeptides, hormones, enzymes in the body according to the physiological requirement. The altered amount of protein in a sample extract may be an indication of abnormality. Also protein concentration determination is important to veryfy the claimed amount of proteins in a sample.

b. No, Different assay method may not have exactly the same result because different method has different sensitivity. For example BSA protein assay which uses absorption technique is considered as best technique for protein determination because it gives more precise and reproducible result.

c. Yes, the relative value may be same if the procedure is appliying properly because the changes occurs between different assay will show relative changes relatively.

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