Yes, we do discourage attachments. Links are better. This is a strong recommendation rather than a hanging offence (we are easy-going people). I've added a sentence to that effect at http://www.ccp4.ac.uk/ccp4bb.php
Regards Martyn On Thu, 2008-01-31 at 08:58 +0000, Frank von Delft wrote: > Actually, it's not book keeping, it's simple courtesy -- and not only on > a BB: an attachment is lazy, and a large attachment is downright rude. > > I am routinely stuck with a slow connection (travelling), and others are > *permanently* stuck with one. So please be nice... ;) > > phx. > > > > Anastassis Perrakis wrote: > > Dear all - > > > > Sorry to intervene on a 'book keeping' issue, but indeed over the last > > few months an increasing number of people (Jerry is not the first, so > > Jerry please do not take it personally) attach pictures etc. I think > > in a bb standard practice dictates to only use text - if illustrations > > are needed to explain the problem, you can put them in eg a web site. > > > > Some text like that was in the 'code of conduct' off ccp4bb in the > > past, but I could no longer find it. > > > > Thus apologies if I am wrong and policies have changed, but maybe the > > ccp4 crowd could tell us what is the suggested policy. > > > > And, if you really want to send an image please do bother to make it > > small. The initial posting had a 630k image, which it took me 1 min to > > make 20k and it still makes the point (attached so I can also violate > > the rules i am suggesting - I love inconsistency). > > > > Thanks, Tassos > > > > > > > > > > On Jan 30, 2008, at 20:11, Jerry McCully wrote: > > > >> > >> Dear All: > >> > >> Thanks a lot for the prompt reply on this topic of salt > >> sensitive complex. > >> > >> Attached please find one ITC final figure done under 25mM > >> Tris(pH8.0), 60mM NaCl. > >> > >> As mentioned before, the ionization of Tris will interfere with > >> the ITC experiments. > >> > >> Therefore I am sure of my binding results. > >> > >> Can anyone give me some comments on this ITC experiment? > >> Basically do these two proteins bind to each other? If so, how should > >> I improve the ITC experiments to get a similar affinity shown by > >> BIAcore(about 0.5uM)? > >> > >> Thanks again. > >> > >> Jerry > >> > >> > >> ------------------------------------------------------------------------ > >> > >> Hi Jerry, > >> > >> Tris can cause problems, you are better off using something like > >> HEPES, and HEPES should be ok at pH 8. (Buffers with an > >> ethane-sulphonic acid group tend to be the best - those ending in > >> 'ES', so MES, TES and HEPES) > >> > >> FYI, the error on your K is bigger than the actual measurement - > >> 1.49x10^5 ± 1.5x10^5. > >> Signal to noise to is probably your enemy, which is making the curve > >> fitting difficult. Changing buffer may help this - there may be some > >> non-specific component to what you're seeing - increasing salt a bit > >> or dropping in something like 5% glycerol may help with this. > >> > >> Would you be able to post a jpeg/pdf of the curve? > >> > >> Regards, > >> > >> David > >> > >> > >> On 25/01/2008, Jerry McCully wrote: > >> > > >> > Dear All: > >> > > >> > Firstly I would like to thank many folks here for giving me > >> great > >> > ideas several days ago. > >> > > >> > The following are some updates for this question. > >> > > >> > I did ITC experiments again using 25mMTris(pH8), 60mM NaCl(low > >> salt > >> > condition). > >> > > >> > But things still turn out to be a little weird. > >> > > >> > I increased the concentration of both proteins(60uM in the cell > >> and > >> > 1200uM in the syringe). At the end of the ITC, I saw a little of > >> > precipitation of both the proteins. > >> > > >> > Fortunately I can roughly fit the curve this time. However, the > >> heat was > >> > still low, around 1Kcal/mole of per injectant. I am not sure about > >> the > >> > fitting statistics. > >> > > >> > > >> > > >> > N 1.10 ±0.17 > >> > > >> > K 1.49E5 ±1.5E5 > >> > > >> > DH -893.5 ±213 > >> > > >> > DS 20.7 > >> > > >> > Was the enthalpy was offset by the ionization of Tris buffer? > >> > > >> > Can I use Hepes buffer around pH8 to do ITC? > >> > > >> > > >> > Welcome any comments about the statistics and suggestions on how to > >> > improve the ITC experiments.have a nice weekend. > >> > > >> > Jerry > >> > > >> > >> > >> > >> > >> ------------------------------------------------------------------------ > >> Helping your favorite cause is as easy as instant messaging. You > >> IM, we give. Learn more. > >> <http://im.live.com/Messenger/IM/Home/?source=text_hotmail_join> > >> > >> > >> ------------------------------------------------------------------------ > >> Shed those extra pounds with MSN and The Biggest Loser! Learn more. > >> <http://biggestloser.msn.com/> > >> <test-ITC-012608.JPG> > > > -- *********************************************************************** * * * Dr. Martyn Winn * * * * STFC Daresbury Laboratory, Daresbury, Warrington, WA4 4AD, U.K. * * Tel: +44 1925 603455 E-mail: [EMAIL PROTECTED] * * Fax: +44 1925 603825 Skype name: martyn.winn * * URL: http://www.ccp4.ac.uk/martyn/ * ***********************************************************************
