Re: [ccp4bb] RES: [ccp4bb] RES: [ccp4bb] Looking for suggestions with protein expression

precipitating other contaminants at their pI, before the affinity purification steps (by dialysing the lysate). Thanks and Regards, Manjula Ramu <https://www.nimhans.kar.nic.in/> On Fri, Jul 10, 2020 at 11:46 AM Jon Cooper < 488a26d62010-dmarc-requ...@jiscmail.ac.uk> wrote: &g

Re: [ccp4bb] COOT crashes when I merge molecules

I also faced this problem. Manjula On Tue, 11 Jun 2019, 1:48 a.m. Tanner, John J., wrote: > Merging EDO into a structure also crashes in Coot 0.8.9.2 running on > linux. > > > John J. Tanner > Professor of Biochemistry and Chemistry > Department of Biochemistry > University of Missouri > 117 Sc

Re: [ccp4bb] Unable to receive mails

Yes, I also realized the same. On Tue, 30 Apr 2019, 11:21 a.m. Petr Kolenko, wrote: > Dear Manjula, #metoo :-D I think it is just a rare occasion that nobody > has posted anything (except you, of course). ;-) Best regards. > > Petr > > > > > > *From:* CCP4 bulletin

[ccp4bb] Unable to receive mails

Dear team, I am not getting any mails from ccp4bb from 27th Apr. Please help me. Manjula To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1

Re: [ccp4bb] Protein Purification- reg

Hello Amala, Usually Ni-NTA won't have this kind of problem of binding. Probably your protein is have interaction with hexagon his tag which is affecting its affinity towards beads. You can try putting tag in C- terminal end of the protein. On Mon, 13 Aug 2018, 8:02 pm Artem Evdokimov, wrote:

Re: [ccp4bb] Protein precipitation

de salts upto 1 to 5 mM. > > On Tue, May 19, 2015 at 10:19 AM, Manjula Ramu > wrote: > >> Hi all, >> >> I am purifying a basic protein with pI 7.8. I used 50mM tris 8.8, 300 mm >> NaCl, 5% glycerol, 0.1mM PMSFand 5mM beta ME in the buffer and performed >> a

[ccp4bb] Protein precipitation

Hi all, I am purifying a basic protein with pI 7.8. I used 50mM tris 8.8, 300 mm NaCl, 5% glycerol, 0.1mM PMSFand 5mM beta ME in the buffer and performed affinity purification. Eluted with 200mM imidazole. While elution I could see slight turbid in eluted protein (I get pure protein, single ban

Re: [ccp4bb] Tev cleavage

Hi, I have used tev protease for tag cleavage during dialysis. In my case always tev got precipitated not my protein. And cleavage was always complete. I havE checked on the sds - page as wel. On 2 May 2015 23:27, "Giulliana Rangel" wrote: > Dear all, > > I'd like some help about my protein cause

Re: [ccp4bb] Strategies to bring out over-expressed protein from inclusion bodies to soluble fraction!!!

Hi, One more suggestion would be autoinduction. This may not help in all the conditions but still can be checked. Instead of IPTG use 0.2% lactose in media and check for expression at different temperature. Hope it helps. All the best. On 17 Oct 2014 21:55, "Roger Rowlett" wrote: > You've trie

[ccp4bb] How to separate BSA present in Thrombin

Hi all, My protein size is nearly the same size of BSA and it has His tag. If I have to cleave the tag I have to use Thrombin which is having huge amount of BSA in it. Now how will I separate this BSA from my protein. Thanks and Regards, Manjula R Research Scholar Department of Biophysics Nationa

[ccp4bb] is small dialysis tubes reusable??

Hi all, I use Slide-A-Lyzer MINI Dialysis Devices from Thermo for dialysis of small protein volumes. Are they reusable? if so how can we store them?? Thanks and Regards, Manjula R Research Scholar Department of Biophysics National Institute of Mental Health and Neurosciences Bengaluru-29, Karnata