ZYMED Brdu STAINING KIT

ZYMED BrdU STAINING KIT

60oC oven 60 minutes Xylene x 3 5 min each 100% ETOH x 3 2 min each 95% ETOH x 2 2 min each 70% ETOH 2 min

1. Quench in 3% H2O2 in methanol 10 min 2. PBS x 3 2 min each o o 3. Trypsin .0625% in dH2O – preheat to 37 C 5 min 37 4. dH2O x 3 2 min each 5. Denaturing solution (reagent 2) 100ul/slide 30 min 6. PBS x 3 2 min 7. Blocking solution (reagent 3) 100 ul/slide 10 min 8. Biotinylated mouse anti-BrdU (reagent 4) 100ul/slide 60 min 9. PBS x 3 2 min 10. Streptavidin-peroxidase (reagent 5) 100ul 10 min 11. PBS x 3 2 min 12. DAB (Vector) 5-10 min 13. Rinse well with dH2O 14. (Optional) Counterstain with hematoxylin 15. Rinse well with dH2O 16. Dehydrate, clear, mount

Incubations are at room temp in a humidified chamber

Hematoxylin counterstain: 1. Diluted hematoxylin (Anatech normal strength) 3 min 1:10 in 1% glacial acetic acid 2. Dip in H2O 3. Scott’s Tap Water 10 dips 4. dH2O