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Rabbit Anti B-gal in paraffin embedded sections

Streptavidin FITC or TexasRed

1. Deparaffinize 20minX2. Rehydrate slides to ddH₂O
2. Wash in ddH₂O 3min X2.
3. Unmasking:
   Set water bath to 95-99°C and prewarm 200ml ddH₂O.
   Add 1.9 ml Antigen Unmasking Solution (Vector) to 200ml ddH₂O.
   Leave in water bath for 20 min.
4. Wash in ddH₂O 1min X2.
5. Wash in PBS 2min X3.
6. Block non-specific proteins with 10% goat serum for 30-60 min at RT. 1ml serum to 9 ml PBT ( 0.1% Tween 20 in PBS )
7. Incubate with Rabbit anti B-gal Ab 1:200 (Cortex Biochem,Inc
   Catalog# CR7001RP2) in 5% goat serum in PBT O/N at 4°C.
8. Wash slides with PBS 5min X6.
9. Incubate with 1:200 Biotinylated goat anti-rabbit IgG (Vector) 1 hour at RT.
10. Wash slides in PBS for 5minX6.
11. Incubate with Streptavidin FITC 1:200 or TexasRed 1:200
12. Wash in PBS for 5min X6.
13. Cover slides with Vectashild mounting medium.

 

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