Delichon Biotechnology Ltd.
FINLAND

Product number 2121-3 Antiserum to histamine Lot No 2121-C

Immunization Protocol

Immunogen: Histamine coupled to keyhole limpet hemocyanin
Animal species: Rabbit
Immunization: Multiple intradermal injections

Applications and Specificity

Applications: Immunocytochemistry of histamine-containing cells and tissues in different animal species. The optimal dilution depends on the tissue, fixation method and detection system. With immunofluorescence method the optimal dilution is genrally 1:1000 - 1:20 000 (in some cases 1:50 000), with biotin-streptavidin-peroxidase technique it is 1:2000 -1: 500 000. The optimal dilution should be tested in each system with serial dilutions (e.g. 1:1000, 1:2000, 1:5000, 1:10 000, 1:20 000, 1:50 000) for best results. The antiserum gives high-quality staining in tissues of flatworm, molluscs, fish, frog, mouse, rat and man. The antiserum requires fixation of the tissues with 1-ethyl-3(3-dimethylaminopropyl)-carbodiimide (EDAC). Optimal conditions are given on a separate application note (No Applic-2121C). The antiserum is not suitable for samples fixed with aldehydes only.

This antiserum is sold as a reagent for research purposes only. Not for in vitro diagnostic use. It is not licensed or approved for adminstration to humans.

Specificity: The specificity of the antiserum has been tested on nitrocellulose filters containing histamine-protein conjugates, L-histidine-protein conjugates, and L-histidine-containing peptides. In these tests, the antiserum detects specifically histamine conjugated to proteins with EDAC. L-histidine containing peptides were not detected. In sections of rat stomach and brain, the antiserum specifically recognized histamine-containing endocrine cells and neurons, respectively. Preabsorption of the antiserum with a histamine-BSA or histamine-ovalbumin conjugate (1-100 ug/ml, depending on tissue) completely abolished the specific reaction. Conjugates are available from Delichon Biotechnology.

Storage: Undiluted antiserum can be stored at -70 C. Lyophilized antiserum can be reconstituted by adding the given amount of sterile water. Repeated freezing and thawing should be avoided. For practical reasons, we recommend that the users divide the antiserum in aliquots which are kept frozen. Glycerol (50%) can be added to prevent repeated freezing.

References:

Polak J, Kendall PA, Heath CM, Pearse AGE (1972) Carbodiimide fixation for electron microscopy and immunoelectron microscopy. Experientia 28: 368-369.

Panula P, Yang HYT, Costa E (1984) Histamine-containing neurons in the rat hypothalamus. Proc. Natl. Acad. Sci. USA 81: 2572-2576.

Håkanson R, Böttcher G, Ekblad E, Panula P, Simonsson M, Dohlsten M, Hallberg T, Sundler F (1986) Histamine in endocrine cells in the stomach. Histochemistry 86: 5-17.

Panula P, Airaksinen MS, Pirvola U, Kotilainen E (1990) A histamine-containing neuronal system in human brain. Neuroscience 34: 127-132.

Panula P, Häppölä O, Airaksinen MS, Auvinen S, Virkamäki A (1988) Carbodiimide as a tissue fixative in histamine immunohistochemistry and its application in developmental neurobiology. J. Histochem. Cytochem. 36: 259-269.

application in developmental neurobiology. J. Histochem. Cytochem. 36: 259-269.