Can anybody recommend the best I have immunostained brain slices (40 um thick)what antifade kit can you recommend for mountingany kits, any recipesI used prolong antifade..
Hi, There are a few options for you. You can either do the mounting media yourself or buy it. Vector sells one called vectashield, which works well and becomes solid.The home made version is not solid and takes up space in your -20ºC freezer.In a dark container dissolve0.05g p-phenylenediamine in 5 ml PBSadd 45 ml glycerine and add 10 drops of 0.1M carbonate-bicarbomate (pH 7.4). Filter and store in a dark container at -20ºC. Discard when dark brown.Platt JL, Michael AF (1983) Retardation of fading and enhancementof intensity of immunofluorescence by p-phenylenediamine. JHistochem Cytochem 31:840–842http://www.jhc.org/cgi/reprint/31/6/840
Have worked with Vectorshield and found that it took a long time to solidify. I used to use something called Elvanol which our lab made up - had to be heated for a long time to get it to disolve but it was great to work with (thick, so had to clip the end off of a yellow tip to pipette it). It went solid quite quickly, tended not to produce air bubbles and allowed FITC signals to be viewed months afterwards (kept at 4 degrees).
Board Rules | Advertise | Privacy | Mobile Feed
© 2004-2008 Scientist Solutions, All Rights Reserved.Resources from NCBI used on this site.