Dear Ben, It was a single crystal that was mounted. I did use IZIT dye, did not trun blue much. Probably its a TCEp crystal. But I did try a negative control without protein+ligand and rest everything same, the drop was clear!
Thank you Regards Kavya On 2023-02-06 22:49, Ben Bax wrote: > Hi Kavya, More than one crystal? > Epitaxial growth (protein crystal growing on salt crystal or vice versa) > Did you try IZIT? > (https://hamptonresearch.com/product-Izit-Crystal-Dye-33.html). > > I have seen salt and protein crystals in the same drop before now - but not > growing together. > Ben > > On 3 Feb 2023, at 22:15, CRAIG A BINGMAN > <000021371e2fba31-dmarc-requ...@jiscmail.ac.uk> wrote: > > Oxidation products of TCEP crystallize in the presence of zinc ions. These > crystals fluoresce strongly under UV, which seems like a dirty trick, if your > expectation is that only protein crystals can do that. The brightfield white > light image of the crystals is also consistent with not-protein crystals, > because of the high refractive index contrast between the crystals and the > mother liquor. > > FROM: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of kavyashreem > <kavyashr...@instem.res.in> > DATE: Friday, February 3, 2023 at 2:32 AM > TO: CCP4BB@JISCMAIL.AC.UK <CCP4BB@JISCMAIL.AC.UK> > SUBJECT: [ccp4bb] Regarding the diffraction image > > Dear all, > > We crystallized a protein (30kDa) + ligand (by cocrystallization), in the > condition 10%PEG3350, 50mM Zinc acetate. > > Protein was in the buffer 20mM HEPES, 150mM NaCl, 1uM ZnCl2, 4mM TCEP, pH 8. > > Crystal: Crystal: > crystal under UV m > > <b06fc576.png> <e091c7fd.png> <8ef9453e.png> > > When we collected the data at an in-house facility, it looked something like > this: > > <b903961d.png> > > The minimum resolution spot is around 9Ang and maximum ~2.2Ang. > > I have not come across a protein diffraction like this, nor of a salt. When I > ran the gel for the incubated protein (protein+ligand), there was no > degradation. > > Although, I was sure there is some problem with this image I tried > processing, which could not be, But indexing showed a unit cell of 11Ang, > 11Ang, 46Ang in P3. which was quite expected for two of the axes but not the > third. > > Can anyone please shed some light on this diffraction image? > > How can it happen? > > Thank you > > Regards > > Kavya > > ------------------------- > > 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 > > ------------------------- > > 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/
