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Methods
EF5 Procedure
Materials
10x PBS
1000x Poisons
4% Paraformaldehyde
1X PBS
1X ttPBS
10mM EF5
Competed ttPBS
Antibody Carrier
Block
Normal Saline
Hoechst 33342
5-bromo-2’deoxyuridine
Peroxidase Reagents
ELK series Antibodies
In Vivo Use
Peroxidase Reagents
- Peroxidase Reagents
- For peroxidase-based staining it is essential to have final rinses and all reagents mixed in azide-free solutions. These include antibody carrier, PBS, ttPBS.
- 3% hydrogen peroxide (4ml stock 30% +4ml 10xPBS + 32ml deionized water).
- Stain (10µl of 3% Hydrogen peroxide + 40µl of AEC concentrate in DMF + 950µl of acetate buffer pH 5.5).
- If crystals are forming in this solution over a typical period of uses (up to 1hour at RT), increase the volume of acetate buffer.
Comments? Questions? E-mail kochc@mail.med.upenn.edu