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Last updated: 12/07/05 / Benjamin Costantino E63-1 Antibody Staining of Drosophila Larval Salivary
 

Summary: Last updated: 12/07/05 / Benjamin Costantino
E63-1 Antibody Staining of Drosophila Larval Salivary
Glands (Whole Mounts)
RECIPES:
PEM
60mM PIPES
5mM EGTA
5mM MgCl2
pH 6.9
PROTOCOL:
1. Make fresh reagents each time: PEM (50mls good for 4 wells), 4% paraformaldehyde
(0.04g paraformaldehyde into 1mL PEM, heat 70C until dissolved)
2. Dissect salivary glands in PBS or D-PBS and keep in buffer until ready for fixing (no
longer than 15min.)
3. Transfer glands into glass slide with wells containing 200ul of 4% paraformaldehyde
fix for no more than 15min. room temp.
4. Transfer fixed glands into Corning Transwell (Corning #3472) insert containing .1ml
of PEM in insert and .6ml in well (NOTE: this volume will always be used unless
otherwise specified). Wash for 3x5 min., RT, light shaking.
5. Permeabilize with 1ml 0.4% Triton X-100 in PEM 3x10min. RT, light shaking.

  

Source: Andres, Andrew - School of Life Sciences, University of Nevada at Las Vegas

 

Collections: Biology and Medicine