Techniques
Buffers and Solutions
Phosphate Buffered Saline (PBS)
Cricket Hemolymph Anti-coagulent
Cell Counts and Cellular Activity
Total Haemocytes (invertebrate blood smears)
Total Leukocytes (vertebrate blood smears)
Lymphoid tissue mass
Sponge implantation
Bacterial killing assay for blood or plasma using plates
Bacterial killing assay for blood or plasma using a spectrophotometer
Phagocytosis assay
Encapsulation
Circulating Immune Molecules (antibodies, cytokines, etc.)
Antigen specific antibodies (indirect ELISA)
Complement
Bacterial killing assay in vitro for insects
Bacterial survival assay in vivo for insects
Real-time PCR for cytokines
Polymerase Chain Reaction (PCR) and electrophoresis gel
C-reactive protein
Mannose-binding protein
Anti-microbial peptides
Lysozyme-like activity for invertebrates
Phenoloxidase and ProPhenoloxidase
Antigen Dosing and Challenging
General insect immune challenge techniques
Fever and Sickness Behavior
Telemetry (radio and internal)
Food consumption/anorexia
Fever
Swelling and Wound Repair
Antigen Detection
Sandwich ELISA
Blood endotoxin levels and clearance
Antigen and parasite clearance
Plaque reduction neutralization test (PRNT)
Molecular techniques
RNA extraction
cDNA synthesis from RNA (invitrogen: SuperScript III First-Strand Synthesis System)
Cloning kit (Invitrogen: TOPO TA Cloning Kit)
PCR Clean-up (Qiagen: QIAquick PCR Purification Kit)
Please use these protocols freely, but we request that if a publication results from their use, the authors acknowledge this website (www.ecoimmunology.org), the author(s) of the protocol used, and the grant that supports the site (NSF-0947177) in your acknowledgements section. Also, please notify us of your publication so that we may list it on our News page.
Please contact Courtney Coon if you have protocols that you would like to share on this website.
Note: This figure is intended to serve only as a guide to those new to the ecoimmunology field. We expect that experts will disagree with our layout, and hope that they will suggest how and why to make alterations. If such comments are reasonable, changes will be made. Additions are welcome too, especially if they come with detailed protocols! In the end, we intend this figure to be a living one, much like Wikipedia. Where available, protocols are underlined and linked below.
