Protein Disappeareance - (Oct/28/2009 )

Hi all:

I´ve got some problems with my protein. I run an SDS-PAGE gel and I identified my almost pure protein. The following day, in order to follow with the experiment I tried to run a Gel filtration chromatography and I discovered that my protein had disappeared. I measure the concentration with the Nanodrop and there was no protein. Just to test, I run again the protein-loading buffer mixture stored from the last SDS-PAGE and surprinsingly, the protein didn´t appear in the gel. Actually, it was no protein in the gel, it didn´t even appear some shorter fragments. The only step between both electrophoresis was a freezing at -20ºC. We have no mix the tubes names and we are sure of their identity. Could someone give us any advice?

Thank you very much

Nostromo

in the future, don't freeze your protein. not all proteins can withstand freezing.

it may have aggregated and remained at the origin.