Methods Flashcards
Radioimmunoassay (RIA)
Add antibody and radiolabeled hormone to sample
Enzymoimmunoassay (EIA)
Add chromogen-tagged antibody to sample; changes color in presence of hormone
Enzyme Linked Immunosorbent Assay (ELISA)
Add antibody and chromogen-tagged hormone to sample
Immunohistochemisty (IHC)
Add chromogen-tagged antibodies to tissue slice
Binds to hormone/receptor
Autoradiography
Add radiolabeled hormone to tissue slice
Accumulates near receptors
Western Blot
Add antibody of protein of interest to gel electrophoresis to determine if present
In Situ Hybridization
Expose tissue slices to radiolabeled cDNA
qPCR
mRNA version of a Western
Ablation + Replacement
Remove and replace (source of) hormone while observing behavior
Lesion
Electrolytic or Neurotoxic destruction of brain area to identify where hormones are synthesized or where they act
Pharmacological
Cannulation to administer antagonists or agonists in specific areas
Tract tracing
Use tracer to identify neural tracts, anterograde or retrograde
Immediate early gene mapping
Use IHC to identify the location of protein products of first genes activated by stimuli
Neural circuitry
Genetic manipulation
KO mice and conditional KO mice
Gene silencing- RNAi and CRISPRi
Neural activity
Electrophysiological recording
Electrical stimulation
Optogenetics!
