[Histonet] Immuno-fluorescence: maximum time in Ab solution?
|From:||Filipa Pontes de Moraes |
I planning to perform a whole mount double immuno-fluorescence for
vessels and smooth muscle cells ( 1Ab solution: rat anti-mouse
Pecam) (2nd Ab sol: Goat anti-rat Alexa488 together with Cy3
conjugated mouse antiSMA) using mouse embryos. I would like to know
what could be the maximum time of 2ndAb solution? Usually is o/n at
4ūC What could happen if instead of one night the embryos stayed 2
days in 2nd Ab solution?
What is the maximum time the embryos could stay mounted in the slides
without loosing the signal? I am planning to mount them in methyl
salicilate (SIGMA) used as the clearing agent.
I am having problems with this clearing agent Methyl salicilate, it
damage my embryos i just add drop by drop and it is still very
difficult to avoid damaging the E10.5 embryos. Can you advice me
other organic clearing agent? i have seen 1 benzil alcohol : 2
benzil benzoate used as cleaing agent and mounting reagent also is it
Thank You very much
Histonet mailing list
<< Previous Message | Next Message >>