Radioimmunocytochemistry using a tritiated goat anti-rabbit second antibody
Affinity-purified goat anti-rabbit immunoglobulin G (GAR) was conjugated with (/sup 3/H)-propionyl succinimidate and used to localize substance P (SP), enkephalin (ENK), and serotonin immunoreactive sites in the spinal dorsal horn and medulla of the rat and cat. Autoradiographic localization was demonstrated on paraffin, frozen, Vibratome, and 2 micron plastic sections. The latter were obtained from radiolabeled Vibratome sections that were embedded in epoxy resin. The distribution of SP, ENK, and serotonin demonstrated by radioimmunocytochemistry was comparable to that observed on semiadjacent sections using peroxidase-antiperoxidase (PAP) immunocytochemistry. The autoradiograms, however, were generated using primary antibody concentrations up to five times more dilute than concentrations used for the PAP procedure. Indirect radioimmunocytochemistry using a (3H) anti-immunoglobulin G second antibody can be used to localize a variety of monoclonal and polyclonal antisera. It is quantifiable at the light microscopic level and can be potentially used with peroxidase histochemistry to double label immunoreactive structures at the ultrastructural level.
- Research Organization:
- Univ. of California, San Francisco
- OSTI ID:
- 6132073
- Journal Information:
- J. Histochem. Cytochem.; (United States), Vol. 32:7
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
ANTIBODIES
LABELLING
BRAIN
RADIOCHEMISTRY
SPINAL CORD
AUTORADIOGRAPHY
CATS
IMMUNOGLOBULINS
PEROXIDASES
RADIOIMMUNOLOGY
RATS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ANIMALS
BODY
CARBOXYLIC ACIDS
CENTRAL NERVOUS SYSTEM
CHEMISTRY
ENZYMES
GLOBULINS
HETEROCYCLIC ACIDS
HETEROCYCLIC COMPOUNDS
IMMUNOLOGY
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAMMALS
NERVOUS SYSTEM
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
OXIDOREDUCTASES
PORPHYRINS
PROTEINS
RODENTS
VERTEBRATES
551001* - Physiological Systems- Tracer Techniques