[Myotox] SDS study of myotoxin-II

[Borries Demeler]

Thanks, Ana, we will follow that protocol. Bruno, I will simply update
the current LIMS project with this information, so no need to make a new
one. 

Regaring the NMR experiments, Paul, Tony and I discussed to get a
baseline profile for just myotoxin-II before jumping into the nanodisk
work. We will also need to get pure nanodisk with the right lipids 
made and measured by both AUC and NMR so we can have the appropriate
background controls. 

Tony and Paul: Please remind us what solutions of myotoxin you will
need, what buffer, and what volumes, and when you can do the
experiments.

Thanks! -Borries


On Wed, Feb 12, 2020 at 11:08:44PM -0300, Ana Gisele da Costa Neves Ferreira wrote:
> Dear Borries,
> As Amy suggested before, I think the first ideia would be to use the same
> experimental conditions assayed by Matsui (attached paper). For SV-AUC
> experiments, the toxin was dissolved in 20 mM Tris-HCl pH 8.0 and 200 mM
> NaCl at a concentration of 1 mg/mL in the presence or absence of 1% (w/v)
> SDS. SDS titration seems to be a good idea too.
> Regarding NMR, what kind of experiment are you planning ?
> Regards,
> AnaG
> 
> Em qua., 12 de fev. de 2020 às 18:02, Borries Demeler <demeler at gmail.com>
> escreveu:
> 
> > Dear Ana and Bruno,
> > We are gearing up to start the AUC experiment in the presence of SDS.
> > Bruno, could you please go into your LIMS account at:
> >
> >         https://uslims.uleth.ca/uslims3_CCH/login.php
> >
> > and update your LIMS project ("Myotoxin-II at high concentration") and
> > explain in the "Buffer Components" and "Experimental Design" section
> > how we should prepare the buffer and protein, i.e., what SDS
> > concentrations to use, what buffer to use, what protein concentrations
> > to try. My suggestion would be to use a moderate concentration (say, 0.5
> > OD 280 nm), and perhaps do a SDS titration, but want to defer to you
> > or Ana in case you have a better idea.
> >
> > Amy is planning the experiments for the end of this week and it would be
> > good to get this info soon.
> >
> > On another topics:
> > Paul and TOny: We have sufficient new material on hand to run the
> > baseline NMR experiments on Myotoxin now. Please send Amy instructions
> > for what concentration, buffer etc. you will need and we will bring it
> > to you. WHen can we run the NMR experiments?
> >
> > Thanks, -Borries
> > _______________________________________________
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> > Myotox at biophysics.uleth.ca
> > http://demeler7.uleth.ca/mailman/listinfo/myotox
> >
> 
> 
> -- 
> Ana Gisele da C. Neves Ferreira
> Pesquisadora Titular em Saúde Pública
> Chefe do Laboratorio de Toxinologia
> Instituto Oswaldo Cruz - Fiocruz
> Pavilhao Ozorio de Almeida, sala 05
> Av. Brasil, 4365 - Manguinhos
> Rio de Janeiro - RJ
> 21040-900 Brasil
> (+5521) 2562-1381 / 98801-5726


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