[Histonet] Immunohistochemistry with whole mouse embryos

[fmoraes <@t> igc.gulbenkian.pt]

Hi

I am having problems to perform some whole mount stainings protocols in mouse
embryos E10.5:

1) The whole mount staining protocol for PECAM (anti-mouse PECAM monoclonal
antibody MEC13.3-Pharmigen) with mouse embryos E10.5 (10dpc). The color
reaction (using DAB takes 10 min and the embryos are all black) however, after
pos-developing washes I don't have any signal. I think is a question of
penetrance of the Antibody. Do you have some tips to permeabilize the embryos?

2) If I want to do an immunofluorescence protocol with whole mount mouse embryos
E10.5 (using a Cy3 conjugated monoclonal Antibody - red spectrum,
Absorbance:552nm Emission:570nm) and I want to analyse the whole specimen under
a confocal microscope. What should be the mounting medium more appropriated?

Thank You very much

Filipa Moraes

Graduate Student
Gulbenkian Institute of Science
Portugal
+315 214464525