Bradford assay is a commonly used method to quantify (estimate) protein levels. Unfortunately, I was confused by the protocols. I have done the assay before. However, different people appear to use different protocols. I do not know which is right (more accurate). Seniors recommend that I read the manufacturer's protocol which should be optimized (Why did not I think of that?!!). Well, it turns out that everyone is partially right in some parts and partially "wrong" (perhaps they took into consideration certain factors which I did not).
Ok. So after consulting the manufacturer's protocol and verifying it with Current Protocols in Molecular Biology and a certain university's protocol, I have come to a final protocol.
1. Protein standards will range from 0.05mg/ml to 0.5mg/ml BSA (0, 0.05, 0.10, 0.20, 0.30, 0.40, 0.50)
2. Add 10ul of standard/sample into each well.
2. Add 10ul of standard/sample into each well.
3. Add 200ul of 5x diluted Bradford regent.
4. Incubate for at least 5 minutes.
5. Read at 595nm.
6. Perform standard curve every time I do the assay (to account for different incubation time).
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