Does anyone have protocol that works nicely for making lysates of virus infected cells to coat the plates for ELISA?
Thanks walter.
But the reason why I posted the question here was to save myself many hours of literature screening and get quick and hopefully good answer from someone dealing with the same thing I deal.
Anyone?
Yes I need only viral proteins and would be very happy to get rid of as much as possible cell structures and debris. Still I don't want to produce huge amounts of virus by standard protocols and purification, as that would be too expensive.
I want to test my anti MCMV antibodies in ELISA, therefore I need to coat viral proteins (antigens) on my plates. I have tried some protocols for lysates, but I couldn't get reproducible results, although they were positive.
@walter
No problem walter. All suggestions are welcome. We are here to help each other by different ways.
Faruk,
in my opinion if you want to test if MCMV antibodies are reactive for viral proteins you cannot do an ELISA coating MCMC-infected cell lysate. You should do an IFA (immunofluorescent assay) seeding on a microscopy slide infected and uninfected cells at 50% ratios, or you should performe a western blot running on a SDS-PAGE MCMC-infected cell-lysate in SDS-loding buffer. I am not suprire that your ELISAs are not reproducible
this is a my old paper where to find some informations, just one of many that you can find on pub-med
Immunological characterization of Toscana virus proteins.
Di Bonito P, Nicoletti L, Mochi S, Accardi L, Marchi A, Giorgi C.
Arch Virol. 1999;144(10):1947-60.
Thanks Paola!
Can you send me pdf of your paper? I don't have subscription for Archives of Virology.
Daniel
what is the problem with researchgate, in your profile you can upload all your papers,
copyright problems?
"Verba volant, scripta manent"
we are scientists , we have to demostrate our claims. In science, sources of informations are important, an article in pub-med is better than someone who speaks from one end of universe,
do not you think?
cheers
paola
"what is the problem with researchgate, in your profile you can upload all your papers,
copyright problems?"
Yes. Wait for publisher' agents.
Did you heare about Open Access to papers ? Actually, it is allowed to make post peer-review published papers openly available with some restrictions that are far less restrictive than most people suspect (see http://www.sherpa.ac.uk/romeo/) - this is beside the "green road" of open access !
ResearchGate uses this database to check which papers one is allowed to upload to one´s profile page (which is considered to be a researchers homepage) - more in the publication section of your profile page !
@Daniel: Generally, sending one´s own papers to requesting scientist is accepted within the scientific community and is done on a daily base - I am not a lawyer, but I assume that nobody could say something against sending one´s own research results to other people (especially not if you send your last word version-not the publishers PDF) - if sb knows cases, pls let me know. However, it is always good to think about copyright issues and obey the laws - more discussions are in Open Access group, or google for open access, ...
Soenke Bartling wrote:
"ResearchGate uses this database to check which papers one is allowed to upload to one´s profile page (which is considered to be a researchers homepage) - more in the publication section of your profile page !"
It's illusion. ResearchGate can't identify the archiving status. See below for its estimates for several Elsevier journals.
Pre-print: Archiving status unclear
Post-print: Archiving status unclear
Please do not SPAM this subject. I was looking for info on ELISA, not on law and copyrights. I believe that author of paper, besides the publisher, holds rights to his/her work.
Daniel, thanks for your contribution.
Sure it is good when someone can do work for me, exactly the same as when I can do work for someone. That is the idea for coming to this place.
And yes, you do spam. Open new thread about copyright and write there as much as you like.
Guys
Do not be polemical, this square is useful because it allows people to communicate with each other,
can give some shortcuts
but everyone must do their work and study study study.....
in science nothing is free
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