I have recently started working with the Insitu hybridisation for Cd31 in P0 mice in cryosections of 10um.
I don’t see any signal . but worked for Embryonic stage ! I dont know how to change the conditions
I tried with digoxigenin labels. I used riboprobes. Can anyone suggesr me or good protocol or good riboprobes for Cd31 (not the commercial)?
Thank you
Hello Suma Sumathi,
you have to enhance the Dig -signal by an immunohistochemical reaction. In short: after stringent washing H2O2-Pretreatment to block endoginous peroxidas; blocking in 10 % normal Goat-serum for 60 min, Anti-Digoxigenin (Roche Sheep Anti-Dig-POX Fab complex) for 90 min, rinse in TBS, enhancement with biotinylated Tyramide 20 min, rinse in TBS, StrepAvidin CY5 or what ever you like 90 min, rinsing in TBS. After airdrying mount the sections in a Dapi containing mounting medium . To be sure that your protocol is working take a negativ and a positiv control.
GenScript generates cDNA for any gene you need. Buty ou have to lable the probes after linearisation by yourself.
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