https://wiki.tanakalab.org/index.php?title=Cocktail_Protocol_for_Pax7&feed=atom&action=historyCocktail Protocol for Pax7 - Revision history2024-03-29T11:40:40ZRevision history for this page on the wikiMediaWiki 1.35.8https://wiki.tanakalab.org/index.php?title=Cocktail_Protocol_for_Pax7&diff=896&oldid=prevBenjamin.wachtel: Created page with "Cocktail Protocol for Pax7 Cocktail Protocol for Pax7/Cy3 and NeuN/Cy5Fab *10µl NeuN *+ 11.8µl Cy5 *+ 80µl PBS/BSA (0.01%) *30’@RT in alufoil *+ 12µl α-BrdU *30’@RT..."2018-07-26T12:43:18Z<p>Created page with "Cocktail Protocol for Pax7 Cocktail Protocol for Pax7/Cy3 and NeuN/Cy5Fab *10µl NeuN *+ 11.8µl Cy5 *+ 80µl PBS/BSA (0.01%) *30’@RT in alufoil *+ 12µl α-BrdU *30’@RT..."</p>
<p><b>New page</b></p><div>Cocktail Protocol for Pax7<br />
<br />
Cocktail Protocol for Pax7/Cy3 and NeuN/Cy5Fab<br />
<br />
*10µl NeuN<br />
*+ 11.8µl Cy5<br />
*+ 80µl PBS/BSA (0.01%)<br />
*30’@RT in alufoil<br />
*+ 12µl α-BrdU<br />
*30’@RT in alufoil<br />
*+ (400µl TBS-Tween + 10% Goat Serum)<br />
*or (900µl PBS/Triton + 10% Goat Serum)<br />
<br />
Procedure:<br />
<br />
#Wash slides 6x10’with TBS-Tween (PBS/Triton)@RT<br />
#Block for 1hour with TBS/Tween (PBS/Triton) + 10% Goat Serum<br />
#Dilute Pax7 1:200 in TBS/Tween (PBS/Triton) + 10% Goat Serum and centrifuge for 10’@13K<br />
#Add to the slides and incubate O/N@4°C<br />
#Wash slides 5x10’with TBS-Tween (PBS/Triton)@RT<br />
#Dilute Goat α-mouse Cy3Fab 1:200 in TBS/Tween (PBS/Triton) + 10% Goat Serum and centrifuge for 10’@13K<br />
#Add 200µl of this to the slides for 2hours @RT<br />
#Start to prepare the NeuN-Cy5Fab Cocktail<br />
#Wash the slides for at least 3x10’ with TBS-Tween (PBS/Triton)@RT<br />
#Centrifuge the Cocktail for 10’@13K<br />
#Add the Cocktail to the slides and incubate O/N@4°C<br />
#Wash the slides at least 6x10 with TBS-Tween (PBS/Triton)@RT<br />
#Add Hoechst (1µg/ml)<br />
#Wash once with TBS/Tween (PBS/Triton) for 10’<br />
#Mount with mounting media or 50% Glycerol/PBS</div>Benjamin.wachtel