[phenixbb] monomer-dimer

intekhab alam faisaldbg at gmail.com
Mon Aug 9 04:38:45 PDT 2010

Hi everyone
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

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