Re: [ccp4bb] offtopic: related to protein purification

Dear Sonia, Have you passed the protein through Size exclusion column and confirmed that it contains a single oligomer? Do you use fresh protein preparation for ITC? If you have tried the above two, then increase the protein concentration. Checking at diff. pH buffer or temperature can also

Re: [ccp4bb] Thermofluor assay

Hi Theresa, Check if this works in your case http://www.sciencedirect.com/science/article/pii/S0969212608000609 good luck !! On Sat, Apr 12, 2014 at 4:38 PM, Theresa Hsu theresah...@live.com wrote: Dear all Does anyone has experience with Thermofluor assay to find the substrate

Re: [ccp4bb] Very sad news: Guy Dodson

Prof. Guy Dodson He is one of the most wonderful person, I have met in my life. He had this very charming and vibrant personality, with whom one can speak about a very broad range of topics starting from Crystallography ... General Science ... Politics. His memory was just wonderful, I discussed

[ccp4bb] off topic: ITC program error

Hi All Sorry for this off-topic, but I need an urgent help. We have an Origin 7 version of VP-ITC Microcal. I had shut it down 2 days back and switched it on today. When I am trying to open the VPviewer2000 where I can start my run it says that error as missing last inj. I had tried to run as

[ccp4bb] sarkosyl

Hi All, I was wondering if anyone has used N-lauryl sarcosine in the lysis buffer for purifying protein and checked if the protein was functional or structurally okay Please let me know kind regards -- rashmi

Re: [ccp4bb] sarkosyl

they sprayed it into a rat's nose the rat died, they had no choice but to put a skull sign on the MSDS. Zhijie *From:* Rashmi Panigrahi rashmi.panigrah...@gmail.com *Sent:* Thursday, September 20, 2012 5:47 AM *To:* CCP4BB@JISCMAIL.AC.UK *Subject:* [ccp4bb] sarkosyl Hi All, I

Re: [ccp4bb] off topic: ITC or Biacore

Thank you all for your suggestions. In the same way can Biacore work be done at lower temperature?? I want to do both ITC and Biacore for my protein and peptide regards Rashmi On Thu, Aug 9, 2012 at 9:19 PM, Edwin Pozharski epozh...@umaryland.eduwrote: On 08/09/2012 03:55 AM, rashmi panigrahi

[ccp4bb] off topic: protein peptide interaction using DSF

Hi all, Has anyone performed protein peptide interaction experiments using flurophore on a real time PCR machine? Please advice me on how the experiment is performed. Can I use ANS? with regards -- rashmi

[ccp4bb] protease inhibitor

Hi All, Does any one add protease inhibitor to purified protein just before setting trays? If yes what is the typical % that should be added ? regards rashmi

[ccp4bb] precipitation.

Hi all, 1) I have a protein which gives two peaks on the 1ml Histrap column, Has anyone seen this kind of behaviour and does this mean that there are two populations of protein. They are partially seperated. 2) I tried to load the two peaks seperately on the superdex-75pg column. They came out as

[ccp4bb] protein lost on membrane of centricon!!

Hi all, I tried to concentrate my protein using vivaspin 20 10,000 MWCO PES. The protein was in 50mM Hepes pH 7.5, 500mM KCl and 10% glycerol. I lost about 90% of my protein on the membrane of the centricon. Please suggest some way of concentrating this protein. Will concentrating using peg

[ccp4bb] dissolving peptides !

Hello everybody, I have a 16 mer peptide which is predicted to be positively charged alpha helix and has 50% of its sequence hydrophobic. Could any one recommend the best way to dissolve it, and then it can be used for crystallization trials. thanks -- rashmi

[ccp4bb] Setting Microbatch trays !!

Hi, While setting up microbatch trays (under oil crystallization), 1. Has anybody used mineral oil (sigma M8410) and obtained some crystal hits? 2. Has anybody used anything other than Al's oil from Hampton, as Parafin oil from sigma or silicone oil from sigma and obtained some crystal hits?