Noradrenaline antibody

Details for Product No. ABIN6559955
Antigen
Reactivity
Hormone
4
3
3
1
1
Host
Rabbit
11
1
Clonality
Polyclonal
Conjugate
Un-conjugated
3
3
Application
Immunocytochemistry (ICC)
9
8
7
3
2
1
1
1
Options
Immunogen Synthetic noradrenaline conjugated to protein carrier (PC)
Isotype IgG
Specificity Conjugated noradrenaline . Using a conjugate noradrenaline-PC, antibody specificity was performed with an ELISA test by competition experiments with the following compounds: Compounds Cross-reactivity ratio (a) Noradrenaline-G-PC 1 Octopamine-G-PC 1/30 Dopamine-G-PC 1/70 Adrenaline-G-PC 1/180 L-DOPA-G-PC 1/>5,000 Tyramine-G-PC 1/>5,000 Noradrenaline 1/>5,000 (a): Noradrenaline-G-PC concentration / other conjugated catecholamines concentration at half displacement G = Glutaraldehyde
Purification Antiserum previously preabsorbed on protein carriers, and purified
Antigen
Target Type Hormone
Application Notes Example of Immunocytochemistry application Detection of conjugated Noradrenaline in rat brain 1- Perfusion: The rat is anesthetized with sodium Pentobarbital or Nembutal and perfused intracardially through the aorta using a pump with the following solutions: Solution A: cacodylate 0.1M, sodium metabisulfite 10g/l, pH = 6.2 Solution B: cacodylate 0.1M, sodium metabisulfite 10g/l and glutaraldehyde 3-5%, pH = 7.5 2- Post fixation: 15 to 30 min in solution B, then 4 soft washes in Tris 0.05M with sodium metabisulfite 8.5g/l, pH 7.5 (solution C). 3- Tissue sectionning: Cryostat or vibratome sections can be used. 4- Reduction step: Sections are reduced with the solution C containing sodium borohydride (0.1M) for 10 min. Then, the sections are washed 4 times with solution C without sodium borohydride. 5- Application of anti-conjugated Noradrenaline antibodies: The final dilution is 1/1,000 to 1/5,000 in solution C containing triton X100 0.5%, plus 2% of non-specific serum. A dozen of sections can be incubated with 2ml of antibody solution overnight at 4°C. Then, after this period, the sections are washed 3 times (10 min) with solution C. N.B.: Antibodies may be used at a higher dilution. The customer should explore the antibody dilution to reduce the possibility of high background. Note that a substitution in the buffer system as used in our protocol may change the background and the antibody recognition. 6 -PAP procedure: Second antibody: Sections are incubated with 1/100 dilution of goat anti-rabbit in solution C for 3 hours at 20°C or 1 hour at 37°C. Then, they are washed 3 times (10 min) with solution C PAP: Sections are incubated with 1/1000 dilution of rabbit peroxidase anti-peroxidase complex in solution C for 1 hour at 37°C. Then, they are washed 3 times (10 min) with solution C Revelation: Antibody-antigen complexes are revealed using diaminobenzidine (25mg/100ml) (or other chromogen) dissolved in Tris 0.05M and filtrated 0.05% of H2O2 is added. The sections are incubated for 10 min at 20°C. Reaction is stopped by transfering sections in 5ml of Tris 0.05M. Recommended dilutions for Immunohistochemistry (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