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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
4% Paraformaldehyde
- 4% Paraformaldehyde
- Weigh out 16g of 95% paraformaldehyde in prill form (Sigma-Aldrich 441244) into coulter vial with lid.
- Heat 360ml of deionized water in the microwave for 3min.
- In glass jar add about half the hot water.
- Add 40 ml of 10xPBS.
- Add 200µl of 1NnaOH.
- Add magnetic stir bar.
- Add PF Prills and rinse coulter vial with remaining water into the jar.
- Stir solution until clear (heating on hotplate may speed up).
- Vacuum filter the solution.
- Rinse glass jar and return filtered solution to jar.
- Measure the pH using pH meter. Should be between 7.2-7.5.
Comments? Questions? E-mail kochc@mail.med.upenn.edu