I tried sokaing ,high concentration of DMSO is effecting crystal, so I tried other contidtion where 10%DMSO is there. Got 1.9 A data with protein co crystalization with 10mM ligand. But ligand occupancy is not visble. I got the KD value from ITC, with reverse titration. With 10uM ligand in cell and 150uM protein in syringe. .
Can anybody suggest some views.please feel free to share your experiences. On Sat, Apr 24, 2021, 10:49 AM Peat, Tom (Manufacturing, Parkville) <tom.p...@csiro.au> wrote: > Hello Gourab, > > DMSO is not the only possible solvent- there are others to try. > I don't want to sound negative, but 40 micromolar is not a very tight > binding compound. It would also depend on how that binding constant was > measured- how did someone get enough in solution to measure that? > Best of luck, tom > > Tom Peat, PhD > Proteins Group > Biomedical Program, CSIRO > 343 Royal Parade > Parkville, VIC, 3052 > +613 9662 7304 > +614 57 539 419 > tom.p...@csiro.au > > ------------------------------ > *From:* CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of Gourab > Basu Choudhury <goura...@csiriicb.res.in> > *Sent:* Saturday, April 24, 2021 12:46 PM > *To:* CCP4BB@JISCMAIL.AC.UK <CCP4BB@JISCMAIL.AC.UK> > *Subject:* [ccp4bb] Co crystalization with less soluble ligand. > > Hello everyone, > > I am finding it difficult for getting a ligand density inside the protein > as the ligand is very much insoluble. It's only soluble in 100% DMSO. I > tried for co crystalization. Kd value of the ligand is near 40um. Any > suggestion what to do? > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
