Sample volume for reverse phase chromatography - (Aug/08/2008 )
Dear all,
I would have a question regarding sample volume for reverse phase chromatography (RPC) protein purification. Is it true, that "RPC is independent of sample volume. Large volumes of dilute sample can be applied in order to concentrate and separate the sample" (I have read this information in a handbook from one company)?
For example, the column volume is 1ml. It should not be difference, whether I load 10ml of (diluted) protein sample (e.g., 500ug proteins) or 1ml of the same but concentrated protein sample (also 500ug)?
Thanks for your advices.
vic
I would have a question regarding sample volume for reverse phase chromatography (RPC) protein purification. Is it true, that "RPC is independent of sample volume. Large volumes of dilute sample can be applied in order to concentrate and separate the sample" (I have read this information in a handbook from one company)?
For example, the column volume is 1ml. It should not be difference, whether I load 10ml of (diluted) protein sample (e.g., 500ug proteins) or 1ml of the same but concentrated protein sample (also 500ug)?
Thanks for your advices.
vic
Shouldn't be a problem.
No problem
Binding capacity should be the same no matter how large the volume your sample is in. I used to use an RP protocol to concentrate a sample following ion exchange and prior to size exclusion chromatography and it worked very well.