Re: [ccp4bb] co-crystallization

Hi YT- We normally prepare our ligand stocks in DMSO and add this to the protein in 3-fold molar excess. The majority of our ligands are quite insoluble and precipitate when the DMSO concentration decreases upon addition to the protein... so I am not surprised that you are seeing this. I

Re: [ccp4bb] Crystal Optimization

Hi Jobi- You might want to try using drop ratios ---we have had great success with this many times. My favorite "additive screen" is using the Hampton Research Crystal Screen HT as an additive screen. I usually start by adding 5% to the well---this has often yielded good crystals where the tr

Re: [ccp4bb] off-topic replay: glycerol in protein stock solutions

Hi Tom-- We have had good luck crystallizing proteins with 5-10% glycerol. In the majority of these cases, the glycerol was included in the buffer for the final purification step. We have also seen several cases where 1,2-propanediol worked much better than glycerol. Hope this helps! annie

Re: [ccp4bb] Crystal Imaging Instruments?

Chris-- We use the Formulatrix Rock Imager & are quite satisfied with it. Thanks! Annie -Original Message- From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Chris Morris Sent: Tuesday, February 08, 2011 8:21 AM To: CCP4BB@JISCMAIL.AC.UK Subject: [ccp4bb] Crystal I

Re: [ccp4bb] Your suggestions needed: Difficulties in reproducing HT crystallization conditions. Thanks!

Joe- You might want to try your original drop ratio of 0.6 ul protein + 0.5 ul well in your optimizations in the 96- well format used to obtain your initial crystals. HTH! annie From: CCP4 bulletin board [mailto:ccp...@jiscmail.ac.uk] On Behalf Of Joe Sent: Tuesday, November 23, 2010 12:03 PM

Re: [ccp4bb] Help with Optimizing Crystals

Matt- You might want to try heating your protein to get rid of unfolded/improperly folded protein. We have used 37C for 10 min with good success, but a time course at different temperatures is the best way to determine which parameters are optimum for your protein. Heat-chill it on ice-centr

Re: [ccp4bb] crystallization plates and robotics

Engin-- We have used the Cybi-Well robot for daughtering from the deep well masterblocks into our crystallization trays for several years and really like it. It is made by Cy-Bio. HTH! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX

Re: [ccp4bb] crystallization of a macromolecular complex

Hi Jan-- What is the composition of your protein buffer? Thanks! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) annie.m.hass...@gsk.com "Jan Rash" Sent by: "CCP4 bulletin board" 05-Mar-2010 09:02 Please respond

Re: [ccp4bb] An alternative to glycerol in keeping protein soluble?

Serah-- We have used 1,2-propanediol to help with protein solubility. It has been included it in our purification buffers and the final protein buffer in the 3-5% range. HTH! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) annie.m.hass

Re: [ccp4bb] how to improve resolution

and/or the precipitant concentrations. We have also had good luck substituting sodium malonate for the ammonium sulfate in our optimizations. HTH! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) annie.m.hass...@gsk.com "rui&qu

Re: [ccp4bb] trouble with protein storage

Marie-Helene-- We have included 3-5% 1,2-propanediol in the protein buffer (as a substitute for glycerol) to stabilize proteins and have seen improvement in the long term storage in many cases. Hope this helps! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228

Re: [ccp4bb] heavy atom derivative choice

Sebastiano-- http://www.sbg.bio.ic.ac.uk/had/ good table of HA and pH ranges http://www.shapirolab.org/lab-links/had.html links to a variety of HA databases HTH! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) annie.m.hass

Re: [ccp4bb] can I try crystallization in high salt?

! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) annie.m.hass...@gsk.com "rui" Sent by: "CCP4 bulletin board" 29-Jun-2009 22:32 Please respond to "rui" To CCP4BB@JISCMAIL.AC.UK cc Subject [ccp4bb] can I tr

Re: [ccp4bb] How to improve crystal which is twinning?

mize the concentration of the reagent to add to the well. HTH! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) annie.m.hass...@gsk.com "Ho-Leung Ng" Sent by: holeung...@gmail.com 20-May-2009 18:39 To "CCP4 bulleti

Re: [ccp4bb] How to improve crystal which is twinning?

find one of these reagents that improves crystal quality/diffraction, I optimize the concentration for that. You can do this with the Mosquito robot for 96 well formator in the 24 well VDX trays. Hope this helps! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919

Re: [ccp4bb] Na/K Phosphate

Hampton Research has a good chart for preparing Na/K phosphate solutions: http://hamptonresearch.com/documents/product/23-000180.pdf Hope this helps! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) annie.m.hass...@gsk.com

[ccp4bb] research position at Glaxo SmithKline --North Carolina

Preferred Qualifications This position requires a BA or BS in Chemistry, Biochemistry, or other related major with at least 5 years of molecular biology or biochemistry experience, or MS degree in a relevant area plus typically a minimum of 3 years relevant experience. Experience with molecul

Re: [ccp4bb] Jeffamine M-600 pHing recipe

Jacob-- We pH'd Jeffamines with concentrated HCl in the hood, but I do not recall how much HCl we had to add to get to neutral pH. Nasty stuff. You might want to contact Bob Cudney at Hampton Research for details since they sell these reagents. HTH! annie Annie Hassell Glaxo Smith

Re: [ccp4bb] Crystallization incubators to recommend?

Catrine-- We have used the Precision Scientific Model 815 incubators for storing crystallization trays. I am not sure if Precision Scientific is still around, but these incubators have been very reliable for years. HTH! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709

Re: [ccp4bb] OT: Crystallisation compatible detergents

ot; with a battery of 10-12 detergents and look for nice sharp peaks off the analytical size exclusion column. This has given us good starting points and clearly ruled out some detergents. HTH! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483

Re: [ccp4bb] purification of phosphorylated proteins-off topic

ion exchange or SEC chromatography after that. HTH-- annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) [EMAIL PROTECTED] "Satheesh Kumar Palani Nathan" <[EMAIL PROTECTED]> Sent by: "CCP4 bulletin board" 25-Nov-2008 16:

Re: [ccp4bb] How to improve twinning crystal in protein crystallization

You might want to try acetonitrile. annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) [EMAIL PROTECTED] "Heng Chiat Tai" <[EMAIL PROTECTED]> Sent by: "CCP4 bulletin board" 16-Oct-2008 16:15 Please respond t

Re: [ccp4bb] Crystal growing in one direction...need suggestions

reen. I have had some very good results with this lately. You can do this with and without seeding. Hope this helps! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) [EMAIL PROTECTED] "shivesh kumar" <[EMAIL PROTECTED]> Sen

Re: [ccp4bb] Substrate(water insoluble) Soaking

up your crystallization screens. Good Luck! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) [EMAIL PROTECTED] "charliswu" <[EMAIL PROTECTED]> Sent by: "CCP4 bulletin board" 10-Oct-2008 23:17 Please respond to "char

[ccp4bb] foam dewar

Jean-Baptiste-- We switched to these foam dewars this year, & everyone in our lab uses them now. After having one of the old style glass ones explode during a late night synchrotron adventure, these foam dewars are a welcome change! annie Annie Hassell Glaxo Smithkline 5 Moore Drive

Re: [ccp4bb] protein complex crystallisation

lize difficult proteins. Hope this helps! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) [EMAIL PROTECTED] "Ron hudson" <[EMAIL PROTECTED]> Sent by: "CCP4 bulletin board" 20-Sep-2008 10:59 Please respond to "Ro

Re: [ccp4bb] Regarding crystallization robotic system

e imager we use is the Formulatrix Rock Imager. Hope this helps! annie Annie Hassell Glaxo Smithkline 5 Moore Drive RTP, NC 27709 919/483-3228 919/483-0368 (FAX) [EMAIL PROTECTED] "BichitraKumar Biswal" <[EMAIL PROTECTED]> Sent by: "CCP4 bulletin board" 18-Sep-2