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Acetylcholine anticorps (ACH) Primary Antibody

ACH Reactivité: IHC Hôte: Lapin Polyclonal
N° du produit ABIN6560001
Plus shipping costs $45.00
1 mg
local_shipping Destination: Etats-Unis
Envoi sous 6 à 7 jours ouvrables
  • Antigène
    Cet anticorp Acetylcholine est non-conjugé
    Immunohistochemistry (IHC)
    Conjugated choline-glutaric acid . Using a conjugate choline-glutaric acid-Protein, antibody specificity was performed with an ELISA test by competition experiments with the following compounds: Compounds Cross-reactivity ratio (a) Choline-GA-PL 1 Choline-allyl alcohol-G-BSA 1/1.6 Phosphatidylcholine 1/>10,000 Choline 1/>10,000 Acetylcholine 1/>10,000 (a): Choline-GA-BSA concentration /unconjugated or conjugated analogs concentration at half displacement. GA = Glutaric anhydride PL = Poly lysine BSA = Bovine Serum Albumin
    Antiserum previously preabsobed on protein carriers, and purified
    Synthetic choline-glutaric acid conjugated to protein carrier
  • Indications d'application
    Example of Immunocytochemical applications Detection of conjugated choline-glutaric acid 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 fixative solution: glutaraldehyde 0.5M, 2-nitrobenzyl alcohol 0.1M, sodium metabisulfite 2- Post fixation: 2h in 0.5M glutaraldehyde solution (pH 7.5) without the 2-nitrobenzyl alcohol, then 4 soft washes in Tris 0.05M with sodium metabisulfite 10g/l, pH 7.4 (solution B). 3- Tissue sectionning: Cryostat or vibratome sections can be used. The sections were washed 4 times in solution B, and incubated for 1h at 37°C in solution B containing 0.2% triton X100, plus 1% of non specific serum. 4- Application of anti-conjugated Acetylcholine antibody: The final dilution is 1/1000 to 1/5,000 in solution B containing 0,2% triton X100, plus 1% 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 B. N.B.: The antibody may be used at a higher dilution. The customer should explore the further 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. 5- PAP procedure: Second antibody: Sections are incubated with 1/200 dilution of goat anti-rabbit antibodies diluted at 1/100 in solution B for 3 hours at 20°C or 1 hour at 37°C. Then, they are washed 3 times (10 min) with solution B PAP: Sections are incubated with mouse peroxidase/anti-peroxidase complex diluted at 1/500 in solution B for 1 hour at 37°C. Then, they are washed 3 times (10 min) with solution B Revelation: Antibody-antigen complexes are revealed using diaminobenzidine (25mg/100ml) (or other chromogen) dissolved in Tris 0.05M and filtrated 0.3% of nickel ammonium sulfate 0.05% of H2O2 were 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 Immunocytochemistry (1/1000-1/5,000) Recommended dilutions for Western Blot (1/1,000-1/2,000)
    For Research Use only
  • Format
  • Antigène
    Autre désignation
    Classe de substances
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