Tyrosine antibody

Details for Product No. ABIN6559964
Antigen
Reactivity
All Species
4
1
1
1
1
Host
Rabbit
3
2
Clonality
Polyclonal
Conjugate
Un-conjugated
1
1
Application
Immunocytochemistry (ICC)
5
3
3
1
1
1
1
Options
Immunogen Synthetic L.Tyrosine conjugated to protein carrier (PC)
Isotype IgG
Specificity Conjugated L.Tyrosine . Using a conjugate L.Tyrosine-PC, antibody specificity was performed with an ELISA test by competition experiments with the following compounds: Compounds Cross-reactivity ratio (a) L.Tyrosine-G-PC 1 L.Phenylalanine-G-PC 1/6,000 L.Tryptophan-G-PC 1/35,000 L.Tyrosine-GA-PC 1/>50,000 L.Tyrosine (free) 1/>50,000 (a): L.Tyrosine-G-PC concentration / unconjugated or conjugated catecholamine concentration at half displacement G = Glutaraldehyde, GA = Glutaric anhydride
Purification Antiserum previously preabsorbed on protein carriers, and purified
Antigen
Target Type Amino Acid
Application Notes Example of Immunocytochemistry application Detection of conjugated L.Tyrosine in rat brain 1- Perfusion: The rat is anaesthetized 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 L.Tyrosine 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. Gemacbio sells other antibodies to conjugated small molecules raised in rat or mouse: used together, these tools could be helpful for immunocytochemistry double labelling. Recommended dilutions for Western Blot (1/1,000-1/2,000) Recommended dilutions for Immunohistochemistry (1/1,000-1/5,000)
Restrictions For Research Use only
Format Lyophilized
Storage 4 °C