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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
Hoechst 33342
- Hoechst 33342
- Make fresh each week of thaw from frozen stock, keep out of light.
- MW is 561, to make 5mM solution, dissolve @ 2.8mg.ml in normal saline.
- For tissue sections or cells in suspension, aim for final concentration of 20µM which should stain fixed cells immediately. Make via: 80µl of 5mM Hoechst, plus 2.0 ml of 10xPBS + 18ml of deionized water. Dip slides directly into this solution in Coulter vial for 5min, followed by rinse in PBS.
Comments? Questions? E-mail kochc@mail.med.upenn.edu