equal amounts of total protein

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fth1
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equal amounts of total protein

Post by fth1 » Thu Apr 21, 2011 2:21 pm

My task was to take equal amounts of total protein from different tissue samples for ELISA. After protein extraction I quantified my total protein using BCA kit. Then I tried to see whether I really succeed in taking equal amounts of protein. For this purpose I took relevant volume from each sample (volume that contains same protein amounts), dissolved it in the buffer to get equal volume in all the samples and performed protein quantification once again using BCA kit. My results showed the following distribution between the samples:

Average = 265.9 ug/ml
Standard Deviation (SD) = 25.0
Min = 213.2
Max = 332.6

My question is whether such a variation (SD=10% from the average) is good enough to claim that all the samples contain the same amounts of the total protein and if not - which SD is acceptable in such experiments.
Thank you.

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canalon
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Post by canalon » Thu Apr 21, 2011 6:55 pm

One of the thing that you would need to know to give areally good answer would be the SD of the BCA kit. Just take a sample, aliquote multiple (3 or more) times and measure the concentration for all of those. Calculate average and SD and compare to your measures.
Patrick

Science has proof without any certainty. Creationists have certainty without
any proof. (Ashley Montague)

fth1
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Post by fth1 » Sun Apr 24, 2011 5:21 am

Canalon, thank you for your suggestion.

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