PAR-1 Staining Procedure
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<center> <b><font>PAR-1 Staining Procedure</font> </b></center>
<center> <hr /> </center>
- Freeze Fracture to permeabilize embryos
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Fixation
15' in -20 degree 100% MeOH
1 x 10' wash in PBS
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Staining
1 x 60' block in goat serum
1 x 60' primary antibody (rabbit-anti-PAR-1 in 1% BSA, 10% serum in PBS) -- optionally incubated 4 hours @ 16 degrees C
1 x 10' wash in PBS
1 x 10' wash in PBS + 0.5% Tween
1 x 10' wash in PBS
1 x 1 hr secondary antibody (1:00 to 1:200)
1 x 10' wash in PBS
1 x 10' wash in PBS + 0.5% Tween
1 x 10' wash in PBS
Reference: Guo, S. and Kemphues, K., 1995, Cell 81:611-620
