Dear Friends

I am trying to stain whole mount of embryo E9.5 and E10.5 using undermentioned way, but i am unable to get any positive stainning to embryonic vasculature which might be a problem of permiabilization of embryo.
   
  So i would like to enquire about some chemicals and their exact concentrations which can be helpful to solve this issue.
   
  I am using follwoing pattern
   
  5% H2O2 in methanol for 5 hrs
   
  (Primary antibody) Rat antimouse CD31, Cat log No. 550274  1/50
   
  (2ndry antibody) HRP conjugated affinipure goat anti Rat IgG code 112-035-167, 1/100 
  Jakson Immuno Research lab.
   
  Vector DAM, Sk 4100, Vector lab
   
   
  
While

       
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