faisaldbg at gmail.com
Mon Aug 9 04:38:45 PDT 2010
Sorry for some non specific query!!!!!
i am working with a protein that shows a dimer in the crystal structure but
when i tried to figure out that with standard molecular markers in gel
filteration (superdex-200, 24ml column) it turned out to be a monnomer.
Native gel analysis after incubating the protein at 20 degree, 37 degree
showed more dimer at 20 degree celcius as compared to 37. I tried similar
strategy in gel filteration by incubating my protein at various
temperature,where a lot of precipitation was observed at 37 degree celcius
and after removing the precipitates i run the gel filteration that has 0.5
ml higher elution volume as compared to samples incubated at 20 degree
celcius and 4 degree celcius.( Is this significant)
Furthermore i have done some experiments in cold room (4 degree) where the
elution volume is stuck at a point irrespective of the conditions (as Flow
rate, concentration of protein etc) and that is higher than that of the room
temperature by 1 ml.
Standard moleculr weight markers also show higher elution volume in cold
room in comparison to the room temperature by 1 ml.
I will be highly obliged if someone suggest some literature or any otherway
to do gel filtrtaion so that i can clearly resolve this issue. Also let me
know if there is some literature available on effect of temperature on the
elution volume of proteins.
Thanks in advance
LABORATORY OF STRUCTURAL BIOINFORMATICS
KOREA UNIVERSITY, SEOUL
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