Salivary Gland Extracts for Western Blotting
Ben Costantino last updated 10/16/05
A. Required Supplies
- 1x complete DPBS
- Lysis Buffer
- Protease Inhibitor Cocktail (Sigma P-8340)
- 2x lamellae
- dissecting tools
- 500ul microfuge tubes
- microscope slide
B. Dissecting Larvae and Preparing Samples
1. Aseptically transfer ~20mL of complete 1x DPBS into a sterile eye dropper or
2. Fill 3 wells of glass dissecting dish with DPBS to provide a wash for larvae.
3. Remove larvae and place them in the wash to clean food and debris off of them.
4. On a clean microscope slide add 1 drop of DPBS.
5. Dissect animals in your preferred technique.
6. Transfer the glands to a 500ul microfuge tube containing 50ul of DPBS.
7. Continue collecting until you have 10-20 glands.