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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
  • Fixation

    15' in -20 degree 100% MeOH

    1 x 10' wash in PBS

  • 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

 

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