Peptide-Based Drug Design

12 Bulet
6. 5 �L or higher volume Hamilton syringe with adapted needles or ultrafine stainless
steel needles.
7. 2.5-mL Eppendorf tubes to prepare the inoculum.
2.2. Sample Collection
2.2.1. Entire Bodies
1. Liquid nitrogen or dry ice for fast freezing.
2. Mortar to reduce the individuals in powder.
2.2.2. Hemolymph
1. Ice-cold water bath.
2. Refrigerated centrifuge (500–1000g).
3. 0.1–1 mL Hamilton syringes or nanoinjector (Nanoject, Drummond Scientific,
Broomall) used as a glass capillary holder for either hemolymph collection or
experimental infection.
4. Phenylthiourea (PTU, Sigma), stock solution 20 mM in ethanol.
5. 2.5-mL low-protein absorption polypropylene tubes—either Eppendorf or Minisorp
NUNC Immuno tubes.
6. Anticoagulant adapted to the animal model investigated (citrate buffer, carbonate
buffer; modified Alsever solution, etc.).
7. Protease inhibitors: aprotinin or/and phenylmethanesulfonylfluoride (PMSF) as
serine protease inhibitor.
2.2.3. Salivary Glands, Gut Tissue, and Any Other Epithelial Tissue
1. Micro-operative dissection equipment (forceps and microscissors) and a binocular
or an optical microscope.
2. 10 mM phosphate-buffered saline (PBS) pH 7.4.
3. Liquid nitrogen.
2.3. Extraction
1. Phenylthiourea (PTU), stock solution 20 mM in ethanol.
2. Protease inhibitors: aprotinin or/and PMSF as serine protease inhibitor.
3. Trifluoroacetic acid (TFA) high-purity (sequencing) grade (Pierce), acetic acid.
4. Ultrapure water (MilliQTM ).
5. Filter units with 0.8-�m membranes (Millex unit, Millipore).
6. Beakers, ice-cold water bath, a stir bar, and a magnetic stirrer.
7. Refrigerated centrifuge (high speed) with rotors (up to 12,000g) adapted to
Eppendorf and Falcon (15–50 mL) tubes.
8. Ultrasonicator specific for cell lysis and Dounce (maximum 152 �m, minimum
76 �m) and Potter apparatus.