Histamine (HIS) antibody

Details for Product No. ABIN100497
Antigen
Reactivity
Chemical
8
5
3
2
1
Host
Mouse
15
2
1
Clonality
Monoclonal
Conjugate
Un-conjugated
2
1
Application
Immunocytochemistry (ICC)
15
8
6
5
4
4
3
3
1
1
1
Options
Immunogen Synthetic histamine conjugated to protein carrier (Pc)
Isotype IgA, kappa
Specificity Conjugated histamine . Using a conjugate histamine-(Pc), antibody specificity was performed with an ELISA test by competition experiments with the following compounds: Compounds Cross-reactivity ratio (a) Histamine-HD-(Pc) 1 1-Methyl-Histamine-HD-(Pc) 1 Histamine-G-(Pc) 1/> 50,000 Histamine-GA-(Pc) 1/> 50,000 Histidine-HD-(Pc) 1/> 50,000 (a): Histamine-HD- (Pc)
Purification The ascitic fluid was purified by ammonium sulfate precipitation
Antigen
Alternative Name Histamine
Target Type Chemical
Application Notes 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 (1/500- 1/1000) anti-conjugated Histamine antibody. 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). Recommended dilutions for Immunocytochemistry (1/1,000-1/5,000) Recommended dilutions for Western blot (1/1,000-1/2,000)
Restrictions For Research Use only
Format Lyophilized
Storage 4 °C