Glutamate, L- Antikörper

Antigen: Glutamate, L-

Klonalität: Polyklonal

Wirt: Kaninchen

Produktnummer: ABIN165471

Produktbeschreibung

Weitere Bezeichnung: Glutamate, L-,

Immunogen: Polyclonal or monoclonal anti-conjugated L-Glutamate antibodies.

Format: Lyophilized

Isotyp: IgG  (Passende Sekundärantikörper)

Spezifität: Glutamate receptor, Polyclonal or monoclonal anti-conjugated L-Glutamate antibodies. . Using a L.Glutamate conjugate, specificity was performed with an ELISA test by competition experiments with the following compounds: Compounds Cross-reactivity ratio Anti-conjugated L-Glutamate antibody 1 Anti-conjugated GABA antibody 1/ 30 Anti-conjugated D-Aspartate antibody 1/ 300 0,0 0,2 0,4 0,6 0,8 1,0 1,2 0,9 1,2 1,5 1,8 2,1 2,4 Concentration Log of competitor (IgG µg/ml) B/B0 Idiotypic conjugated L.Glutamate response Idiotypic conjugated D.Aspartate response Idiotypic conjugated GABA response Idiotypic conjugated L-Glutamate response inhibition: IC50 = 12µg/ml Idiotypic conjugated GABA response inhibition: IC50 = 400µg/ml Idiotypic conjugated D-Aspartate response inhibition: IC50 = 4,000µg/ml

Anwendungen

Applikationshinweise: Example of immunohistochemistry protocol Perfusion (Example for Adult male Sprague Dawley (weight around 0.5 kg)). 1-The animals can be deeply anaesthetized (for example with urethane-0.5-1.5g/kg, intraperitoneal). 2-Perfused via the ascending aorta with 50 ml of NaCl 9g/l (Heparinized) and pass trhough the system 800-1000 ml of cold 4% paraformaldehyde (Merck) in 0.1 M PB, pH 7.2-7.4, (ten minutes). 3-Dissect out the organs and place in a solution of 4% paraformaldehyde in 0.1M PB, pH 7.2, at 4ºC for twelve to sixteen hours. Immunohistochemistry 1-In order to avoid possible interference with endogenous peroxidase, free-floating sections will be treated with distilled water containing NH 3 (20%), H 2 O 2 (30%) and NaOH (1%) for 20 min (other method is using a solution with 33% of H 2 O 2 and 66% of methanol). 2-Then, wash the sections for 20 min in 0.15 M phosphate-buffered saline (PBS) (pH 7.2) 3-Pre-incubate for 30 min in PBS containing 2-10% (variable to adjust) of normal horse serum and 0.3% of Triton X-100 (mixed solution). 4-Incubate at room temperature (1h 30min) and overnight at 4º C in the same mixed solution containing the diluted anti-anti conjugated L.Glutamate antibodies(1/1,000-1/5,000). 5-Then, the sections will be wash in PBS (30 min). 6-After that we will incubate for 60 min at room temperature with biotinylated anti-(species) immunogammaglobulin (Vector) diluted 1/200 in PBS. 7-Wash during 30 min with PBS. 8-Sections will be incubated for 1 h with a 1/100 diluted avidin-biotin-peroxidase complex (Vectastain) in the mixed solution. 9-After that we will wash the sections in PBS (30 min) 10-Wash with Tris-HCl buffer (pH 7.6)(10 min). 11-The tissue-bound peroxidase will be developed with H 2 O 2 using 3, 3' diaminobenzidine as chromogen. 12-Finally the sections will be rinsed with PBS and coverslipped with PBS/Glycerol (1/1).

Lagerung: After reconstitution with 50µl of distilled water and 50µl of glycerol, the aliquot can be repeated by frozen at -20°C (up to five times).

Beschränkungen: Nur für Forschungszwecke einsetzbar

Publikationen

Publikationen: Jennings, Cotton: "Structural similarities among enzyme pterin binding sites as demonstrated by a monoclonal anti-idiotypic antibody." in: The Journal of biological chemistry, Vol. 265, Issue 4, pp. 1885-9, 1990 (PubMed).

Bona: "Parallel sets and the internal image of antigen within the idiotypic network." in: Federation proceedings, Vol. 43, Issue 10, pp. 2558-62, 1984 (PubMed).

Pillet, Paon, Vorobiev et al.: "Idiotypic network mimicry and antibody catalysis: lessons for the elicitation of efficient anti-idiotypic protease antibodies." in: Journal of immunological methods, Vol. 269, Issue 1-2, pp. 5-12, 2002 (PubMed).