Re: [ccp4bb] crystallization incubators

2008-06-23 Thread Demetres D. Leonidas
Dear Alexandra, we are using the VELP FOC incubators (http://www.velp.com/en/product/5/Environment_line/101/Refrigerated_incubators/143/FOC_225I/) around 2000 Euros. Demetres Alexandra Deaconescu wrote: Hello everyone: I know this question has been posted before, but what models of

[ccp4bb] ccp4mg: SO4 ions

2008-06-23 Thread PhilEvans
I''ve got a structure with a pile of sulphate ions in the PDB file, all in chain X. However, in the molecule selection window, these sulphates do not appear as monomers, nor does chain X appear in the chain list: should they not be there? I can select them explicitly as //X/*, but I'm

Re: [ccp4bb] B-factor problem

2008-06-23 Thread Eleanor Dodson
Sampath Natarajan wrote: Dear All, I am refining a structure with 2.5A resolution by refmac5. I could find the solution by MR using molrep. After fitting the model, I refined the structure again with 0.3 weighting term, but the output PDB file shows many splits in the residues. So I

Re: [ccp4bb] re-indexing P 4 to P P4 21 2

2008-06-23 Thread Eleanor Dodson
Roni Gordon wrote: I have used the CCP4 program re-index to add a screw axis before (e.g. P6 - P61, P2 - P21), but never to change the point group (e.g. PG4 - PG422). I initially re-indexing the reflections without error, but refmac5 later complained that there were too many reflections for the

Re: [ccp4bb] Question about map cutting..

2008-06-23 Thread Eleanor Dodson
Partha Chakrabarti wrote: Hi, Is it possible to cut out a spherical map around a heavy atom site directly without having to make bones or building something? I want to place that in a larger P1 cell and use that for phased molecular replacement (Phaser / Molrep).. can someone point me how to do

[ccp4bb] Econo column with AKTA FPLC

2008-06-23 Thread Exec
Dear All, I have econo column from Biorad and AKTA FPLC system. how shall i connect the econo column to AKTA FPLC system i.e what are the different connections / tubings to be used since akta system the tubing are all small sized and econo column tubing are large capacity. does anyone have

[ccp4bb] Sephadex resins

2008-06-23 Thread Exec
does anyone have protocols / method of sephadex resin. about swelling the resin use of buffers, elution of sample from the resin. i have referred to the GE healthcare website, it gives details of other resins well, but sephadex resin not much detail is given. kindly guide

Re: [ccp4bb] Econo column with AKTA FPLC

2008-06-23 Thread artem
What you need is a tubing and connector kit. Every decent lab has one, and you also usually can order these in part or whole from FPLC/HPLC manufacturers. The two types of female fittings are then attached to a short length of tubing and presto. Artem Dear All, I have econo column from Biorad

Re: [ccp4bb] Sephadex resins

2008-06-23 Thread artem
Any basic chromatography (for proteins) text will have those. It's generally not hard - if you have dry sephadex just resuspend it in 10-20% alcohol, stir well (but gently!) and let sit overnight, then pack the column, wash, and go. Artem does anyone have protocols / method of sephadex resin.

[ccp4bb] REMINDER: Ninth International School on the Crystallography of Biological Macromolecules

2008-06-23 Thread Derek Logan
*** REMINDER *** REGISTRATION IS OPEN for the Ninth International School on the Crystallography of Biological Macromolecules Como (Italy), 29th September – 3rd October 2008 for more details and to register, please visit the web page: www.crystallographyschool.org DEADLINES Registration:

Re: [ccp4bb] re-indexing P 4 to P P4 21 2

2008-06-23 Thread Roni Gordon
Perhaps I should have mentioned that I'm working with an MTZ file created from merged scalepack input... I was simply wondering I can't seem to use sortmtz to perform the k-h, h-k, -l-l transformation and have it merge these directly? It doesn't seem to want to merge from a merged file... Roni

Re: [ccp4bb] Sephadex resins

2008-06-23 Thread R.M. Garavito
The primary reason they don't have much on the Sephadex resins is that the are quite old-fashioned (and also relatively cheaper than most of their high-end resins). However, old Pharmacia/LKB had some really great manuals on chromatography that are still quite useful for new biochemists.

Re: [ccp4bb] query on DNA-protein complex preparation for crystallization

2008-06-23 Thread Phoebe Rice
You should add some salt when you anneal!!! The duplex is highly negatively charged, so adding even a small amount (like 10mM NaCl) will help with charge screening, thus making the two strands less repellant to each other. Buffer is also always a good idea. At low pH and high temp you'll

Re: [ccp4bb] inclusion bodies

2008-06-23 Thread Arundhati Chattopadhyay
You can coexpress the enzyme with chaperone (e.g GroEL and GroES)very often this is very successful. (There are bunches of ways though...) Best wishes, Arundhati 2008/6/23 yangliuqing [EMAIL PROTECTED]: Hello,everyone! Sorry to bother you! Now I have an enzyme cloned in pQE2

Re: [ccp4bb] Econo column with AKTA FPLC

2008-06-23 Thread Nathaniel Echols
On Mon, Jun 23, 2008 at 4:09 AM, Exec [EMAIL PROTECTED] wrote: Dear All, I have econo column from Biorad and AKTA FPLC system. how shall i connect the econo column to AKTA FPLC system i.e what are the different connections / tubings to be used since akta system the tubing are all small

[ccp4bb] RI and MALS for membrane proteins

2008-06-23 Thread John Luz
Would like to know what experiences people have had using in line refractive index and multiangle light scattering detectors for membrane protein purification. Specifically, if the information gathered from such experiments has been useful in optimizing solution conditions for membrane protein

Re: [ccp4bb] Fix hydrogen in PDB and use of reduce

2008-06-23 Thread Bryan Arendall
Reduce adds hydrogens to HETATMs according to its hetdict, which is a text file in the PDB Format for the chemical component dictionaries. A dictionary file is available for download from the url that you cited. Reduce will find the hetdict from either it's compiled path location, the location

Re: [ccp4bb] RI and MALS for membrane proteins

2008-06-23 Thread Brandao-Neto, J (Jose)
dear john, the reference for this in my opinion is still the 1994 paper by george and wilson where they describe the use of the second virial coefficient parameter as a metric for quantifying the potential that certain conditions have to favour crystallisation. see for instance bonete and

[ccp4bb] about anisotrophic diffraction

2008-06-23 Thread Ji lee
Dear, I have a crystal diffracted anisotrophically. I tested with a few different cryo conditions like oil, glycerol in different concentration to get a better data but these conditions didn't help any. Using capillary method improved the diffraction (isotrophic) but the crystal couldn't survive

Re: [ccp4bb] about anisotrophic diffraction

2008-06-23 Thread Jacqueline Vitali
Look at the paper (search in pubmed) by B. M. Schick and F. Jurnak on cryoprotection of TuTs. He changes the reservoir in 24 steps using a 24 well cryschem plate. There are some references there that are important too. Jackie Vitali On Mon, Jun 23, 2008 at 8:14 PM, Ji lee [EMAIL PROTECTED]