Re: [ccp4bb] Micro/Macro crystal seeding experience

Wholeheartedly agree on all these points! Works wonders. -- Kelvin Lau https://people.epfl.ch/kelvin.lau Protein production and structure core facility - PTPSP EPFL SV PTECH PTPSP AI 2146 (Bâtiment AI) Station 19 CH-1015 Lausanne Switzerland Email: kelvin@epfl.ch

[ccp4bb] RES: [ccp4bb] Micro/Macro crystal seeding experience

xta-feira, 18 de dezembro de 2020 12:50 Para: Rafael Marques<mailto:rafael_mmsi...@hotmail.com>; CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK> Assunto: Re: [ccp4bb] Micro/Macro crystal seeding experience We recently had a projects where removing the HIS-tag changed an unsolvable

Re: [ccp4bb] Micro/Macro crystal seeding experience

We recently had a projects where removing the HIS-tag changed an unsolvable crystal form into one where Molecular Replacement worked immediately (and with an NMR ensemble as the search model to boot). In this case the HIS-tag was about 5% of the total domain mass and the tags were probably tangled

Re: [ccp4bb] Micro/Macro crystal seeding experience

<mailto:scott.wa...@nih.gov> https://ccr.cancer.gov/Chemical-Biology-Laboratory/scott-walsh?qt-staff_profile_tabs=3#qt-staff_profile_tabs From: Patrick Shaw Stewart Reply-To: Patrick Shaw Stewart Date: Friday, December 18, 2020 at 5:49 AM To: "CCP4BB@JISCMAIL.AC.UK" Subject: Re: [ccp

Re: [ccp4bb] Micro/Macro crystal seeding experience

Hi Rafael Just to amplify Matthew and David's point about random microseed matrix screening - 1. The idea is you add seeds to RANDOM SCREENS - not everyone understands this! People find it very hard to "let go" of the first conditions they see that give them crystals. 2. I now have the

Re: [ccp4bb] Micro/Macro crystal seeding experience

: [ccp4bb] Micro/Macro crystal seeding experience Hi Raphael Think about simple refinement, differents kits are available additifs kits: - additif, Gras and Silver bullet : from Hampton research - Angstrom and Morpheus from MDL - JBScreen Plus from Jena bioscience detergent kits

Re: [ccp4bb] Micro/Macro crystal seeding experience

Hi Raphael Think about simple refinement, differents kits are available additifs kits: - additif, Gras and Silver bullet : from Hampton research - Angstrom and Morpheus from MDL - JBScreen Plus from Jena bioscience detergent kits: - from Hampton resaerch - from Jena

Re: [ccp4bb] Micro/Macro crystal seeding experience

I want to second the recommendation to try microseed matrix screening. I recently had a case of a protein that did not yield any crystals after trying more than 500 conditions. Of those 500, one single condition gave to me what appeared to be crystalline material, but not distinct single

Re: [ccp4bb] Micro/Macro crystal seeding experience

Hi Rafael, have you considered crystal dehydration? https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3317743/ It’s not easy to implement, but high solvent content could be a reason why your crystals don’t diffract beyond 4A. Did you manage to