View source for Labeled AB staining

<h4>Pax7  + cy3 + NeuN – cy5Fab</h4>

<h3>NeuN – cy5Fab cocktail for 2 slides</h3>
<li>10 µl NeuN (0.5 mg/ml)<li>
<li>11,8 µl cy5 (0.85 mg/ml)<li>
<li>80 µl PBS/ BSA (1mg/ml BSA in PBS → sterile filter)</li>
<li>30 min @ RT in alu foil</li>
<li>12 µl α - BrdU → 30 min @ RT in alu foil</li>
<li>400 µl TBS – Tween + 10% goat serum</li>
<h3>Procedure:</h3>
# Wash slides 6 times 10 min with TBS – Tween @ RT
# Block for 1 h with TBS – Tween + 10% goat serum
# Dilute Pax7 1:200 in TBS – Tween + 10% goat serum and centrifuge for 10 min @ 13000 rpm
# Add to the slides and incubate o/n @ 4 degree
# Wash slides 5 times 10 min with TBS – Tween @ RT
# Dilute goat α - mouse cy3Fab 1:200 and centrifuge for 10 min @ 13000 rpm
# Add 200 µl of goat α - mouse cy3Fab to the slides for 2 h @ RT
# Start to prepare the NeuN – cy5Fab cocktail
# Wash the slides for at least 3 times 10 min with TBS – Tween @ RT
# Centrifuge the cocktail for 10 min @ 13000 rpm
# Add the cocktail to the slides and incubate o/n @ +4 degree
# Wash the slides at least 6 times for 10 min with TBS – Tween @ RT
# Hoechst (1µg/ml) for 10 min @ RT
# Wash once with TBS – Tween for 10 min @ RT
# Mount and check