Dear All
I am purifying a protein from liver. In first step I spotted protein in Flow
Through and could see intense band in SDS PAGE. I pooled the fraction and did
second column. Surprisingly I could not see UV absorbance or even any band in
the SDS. I used PMSF as protease inhibitor from
Dear All
I am running ITC experiment for my protein. At 250 nm Ligand concentration
(protein was 40nm) I was getting exothermic thermogram. But it was not
saturated. I reduced Ligand concentration to 15 nm and checked again. I got
endothermic thermogram. I dont understand why I am getting both
Dear All,
This is an off-topic question. I have protein solution of heterogeneous
(contains both monomer and dimer). I want to perform ITC with this protein. I
doubt whether this heterogeneity will interfere the binding study.
Any advice please.
Thank you
Sajid
Hi all
If anyone have, please post the calibration profile for GE Sephacryl S100 26/60
Hiprep column. Sorry I could not locate in the GE Healthcare site.
Thankx in advance
Sajid
Dear All
Can any one mention some reference regarding the binding of NADPH or ADP in
the surface of the protein. Is there any rules that dominate these two
cofactors to select the surface istead of typical Rossmann fold.
Thank you
Sajid
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Hi All
I have some density and I can fit a peg molecule in their. I am confused and
when I search database for peg3350, I have no hit. How can I build a model for
Peg 3350. I found some PDB's with peg; but they they have used peg400; Is there
any difference in the model between peg3350 and
Dear All
Is there any easy to way write topology and parameter file for a cofactor with
specific conformation.
Thanks a lot
Sajid
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Hi all
I am trying to run autoSHARP. When I submit the job I got following message and
the job is not running. Does any one have got error like this when you start
autoSHARP
Any advice please
Sajid
**
(In case of
Dear All
I want to run MrBump. We have dynamic internet system in our institute. So I am
not able to locate the proxy address.
Any help please
Sajid
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Hi All
I have a data set of Se-Met crystal 3.65A resolution collected at Inflection
and peak wavelength. Also I have a data set at same resolution in native form.
The protein size is 28000 dalton. I dont have any model for this protein. So I
started to find the heavy atom peak search to find
Dear All
Thanks for your suggestions to run revise. Suggestions
from Mike Latchem helped me to overcome the problem.
I have another question.
I am trying to run REFMAC5. It is giving some warning
message.
#
There is no file name for parameter MAPOUT1
The CCP4 program is
Hi All
Sorry for non CCP4 question. I want to make Pottasium Nitrate solution of pH
6.9 (ofcourse for crystallisation trails only). I'm adjusting pH by using NaOH.
But it seems the pH is not stable after sometime. Is there any good way to do
it.
Thank you
Sajid
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Hi All
I'm getting clusters of crystals (tiny and long needles) and I'm trying to get
good single crystals. Even I'm getting the same result at low salt
concentrations. I reduced the temperature to slow down the evaporation, still
no change. So I'm trying with mineral oil. Here I'm getting
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