Re: [ccp4bb] how to purify protein in its native form

Hi: I have seen some protocols(IMPACT from New England Lab etc.) which can overexpress protein with a tag then cut or displace the tag off to get a native protein. Maybe those are the options. Best, Hongnan Cao UCR __

[ccp4bb] problem with cns compile

Ok, I have tried sending this to the cns board and it has been reject with my old email and new email addresses so I will try here and sorry for the non ccp4 question. While compiling CNS 1.21 on an older systems running Redhat Enterp. 4 the following error comes up. machine_f.o(.text+0x11ca

Re: [ccp4bb] how to purify protein in its native form

Yes, However the even holier single method suitable for purification of proteins from native sources is immuno-capture. If you have a good antibody (i.e. an antibody that recognizes a native epitope) you can get very high purity very quickly. If you don't have an antibody - you can get polyclonals

Re: [ccp4bb] how to purify protein in its native form

The holy trinity of protein chromatography is ion exchange (IEX), hydrophobic interaction chromatography (HIC), and gel exclusion (GEC), assuming no affinity purification is possible. Depending on the abundance of protein, these three steps, perhaps in concert with additional chemical steps, in

[ccp4bb] Postdoctoral Fellow: Protein expression and X-ray structural analysis at ZMBP, Tuebingen

Posted on behalf of ZMPB: # There is an opening for a Postdoctoral Student (Protein expression and X-ray structural analysis) in the Department of Plant Biochemistry at the Center for Plant Molecular Biology (ZMB

Re: [ccp4bb] how to purify protein in its native form

Hello Jerry, I would be glad to provide protocol suggestions, but it'd help to know a bit more regarding what you are working on. If you're planning to purify material from its native source (i.e. someone's chopped up sittin' muscle?) - this would be tremendously different from purifying protein f

[ccp4bb] Calculation of angle between two helix of different subunits

Hello all I am interested to know about any programme which can calculate the angle between the helices of different subunit not the consecutive helices. I know about helixang which is a ccp4 supported programme but, i am not sure that does it calculate the relative angle between the two helix of

[ccp4bb] how to purify protein in its native form

Hi, All: Although it is off-topic, definitely I think I can get some help here because we crystallographers are dealing with protein purification almost every day. My protein was expressed in E.coli as a soluble octamer with or without his-tag revealed by gel-filtration. For the

Re: [ccp4bb] DM and MR

I have to use refmac for licencing reasons, sadly, unless someone can recommend an entirely free refinement program to anyone including industry/people in limbo etc. Actually, I was just thinking. In the new MTZ files (which helpfully I dont have on me) there are new values with 'DM' added to the

Re: [ccp4bb] DM and MR

..and I may add that for DMMULTI two "rather nonisomorphous" data sets of the same crystal form can work like a dream, in particular at low resolution where solvent flattening and histogram matching can be tricky, see for example Morth et al. 2007, Nature 450, 1043 & Pedersen et al. 200

Re: [ccp4bb] DM and MR

Dear James and Andy, my twopenny worth. I am using dm and dmmulti to solvent flatten after molecular replacement phases when no shred of experimental phases is available. If the model is severely incomplete (say at least 25% missing or more) I use phases fro

Re: [ccp4bb] DM and MR

Your split between Rfree and R suggests you are using refmac. (I also got a hint when you explicitly said you were using refmac.) Although there are many efficient ways to get stuck in a refinement, I have found the most effective by far is underweighting the structure factors and falling i

Re: [ccp4bb] DM and MR

Jusht my two penneth... Any list of DM programs worth a punt after MR *musht* *shurely* include *resholve*? Resolve ± 'prime&switch' ± NCS pretty much always comes up with the goods for me. Dave 2009/4/15 James Stroud : > Hello All, > > What is the current state of the art with respect to densit

Re: [ccp4bb] DM and MR

Hi James, I usually use DM in teh CCP4 suite to do DM. It gives you options to do solvent flattening, Histogram matching and NCS averaging. If you compare the FOM to the DMFOM in the mtz files, you can usually see and improvement in the score. That is usually all I bother my arse to look at.

[ccp4bb] DM and MR

Hello All, What is the current state of the art with respect to density modification after molecular replacement? In other words, what programs are people having the best luck with these days? Thanks in advance for any hints. James