AB5522 Anti-Acetylcholine Antibody

AB5522
100 µL  
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      Overview

      Replacement Information

      Key Specifications Table

      Species ReactivityKey ApplicationsHostFormatAntibody Type
      R IHC Rb Serum Polyclonal Antibody
      Description
      Catalogue NumberAB5522
      Brand Family Chemicon®
      Trade Name
      • Chemicon
      DescriptionAnti-Acetylcholine Antibody
      References
      Product Information
      FormatSerum
      PresentationRabbit antiserum. Liquid with 0.05% sodium azide.
      Quality LevelMQ100
      Applications
      ApplicationAnti-Acetylcholine Antibody is an antibody against Acetylcholine for use in IH.
      Key Applications
      • Immunohistochemistry
      Application NotesImmunohistochemistry: 1:250-1:2500 by PAP (see suggested protocol below).

      Optimal working dilutions must be determined by the end user.

      PROTOCOL for Acetylcholine Detection by Immunohisto/cytochemistry. Example for a rat brain.

      1. SOLUTIONS TO BE PREPARED - Solution must be prepared as needed.

      Solution A: 0.5M glutaraldehyde, 0.1M 2-nitrobenzyl alcohol, 10g/L sodium metabisulfite, 0.1 M cacodylate, 0.01M sodium bromide, pH 10.7.

      Sometimes a precipitate may form while making solution A. To avoid this, first prepare a mixture with 1.0M glutaraldehyde and 0.2M 2-nitrobenzyl alcohol. Prepare a second mixture with 20g/L sodium metabisulfite and 0.2M cacodylate; add sodium bromide to obtain pH 10.7. Then combine the two mixtures in equal parts while stirring.



      Solution B: 0.05 M Tris, 10.0 g/L sodium metabisulfite, pH 7.4.(*)

      2. RAT PERFUSION - The rat is anaesthetized with sodium pentobarbital or Nembutal and perfused intracardially through the aorta using a pump with Solution A (200-300 mL/min).

      3. POST FIXATION: 2 hours in 0.5M glutaraldehyde solution, (pH 7.5) without the 2-nitrobenzyl alcohol, then 4 soft washes in solution B.

      4. TISSUE SECTIONING: Vibratome or cryostat sections can be used.

      The sections are washed 4 times in solution B, and incubated for 1 hour at 37°C in solution B containing 0.2% Triton X100, plus 1% non specific serum.

      5. APPLICATION OF ACETYLCHOLINE ANTIBODY: Use a final dilution of 1:500-1:2,500 in Solution B containing 0.2% Triton X100 and 1% non-specific serum. Incubate 12 sections per 2 mL diluted antibody overnight, +4°C. Then wash the sections three times for 10 minutes each in Solution B. (Note that the antibody may be usable at a higher dilution. This should be explored to minimize the possibility of high background. Additionally, note that a change in the buffering system as indicated in the protocol may change the background and antibody recognition). The specific reaction is then revealed by PAP procedure.

      6. SECOND ANTIBODY: Incubate the sections with a 1:50 to 1:200 dilution of goat anti-rabbit in Solution B containing 1% non-specific serum for either 3 hrs at 20°C or 2 hr at 37°C. Then wash the sections, 3 times, for 10 minutes each with Solution B.

      7. PAP: Incubate the sections with the appropriate dilution of peroxidase anti-peroxidase (for free floating method) in Solution B containing 1% non-specific serum for 1-2 hours at 37°C. Then wash sections 3 times for 10 min each in solution B.

      8. VISUALIZATION: The antigen-antibody complexes are visualized using DAB-4-HCl (25 mg/100 mL) (or other chromogen) in 0.05M Tris and filtrated; 0.05% hydrogen peroxide is added. Incubate the sections for 10 minutes at room temp. Stop the reaction by transferring the sections to 5 mL 0.05M Tris. Wash tissue with solution D using 2, 10 min washes. Mount sections on chrome-alum coated slides. Dry overnight at 37°C. Rehydrate sections using conventional histological procedures. Coverslip using rapid mounting media.
      Biological Information
      ImmunogenAcetylcholine-Glutaric anhydride-PL
      HostRabbit
      SpecificityAcetylcholine



      The cross-reactivities were determined using either ELISA or RIA techniques, at concentration/unconjugated or conjugated amino acid concentration at half displacement with the following compounds:



      Compound Cross-reactivity

      Choline-GA-PL 1

      Phosphatidylcholine 1/>10,000

      Choline 1/>10,000

      Acetylcholine 1/>10,000

      Choline-allyl alcohol-G-BSA 1/1.6



      The antisera was also tested for specificity using the free-floating PAP technique on rat cortex.



      Abbreviations:



      (GA) Glutaric anhydride

      (=) Non-reduced conjugate

      (PL) Poly lysine

      (BSA) Bovine Serum Albumin
      Species Reactivity
      • Rat
      Antibody TypePolyclonal Antibody
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsMaintain at -20°C in undiluted aliquots for up to 6 months after date of receipt. Avoid repeated freeze/thaw cycles.



      During shipment, small volumes of antibody will occasionally become entrapped in the seal of the product vial. For antibodies with volumes of 200 μl or less, we recommend gently tapping the vial on a hard surface or briefly centrifuging the vial in a tabletop centrifuge to dislodge any liquid in the container's cap.
      Packaging Information
      Material Size100 µL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      Anti-Acetylcholine Antibody SDS

      Title

      Safety Data Sheet (SDS) 

      Anti-Acetylcholine Antibody Certificates of Analysis

      TitleLot Number
      RABBIT ANTI-ACETYLCHOLINE POLYCLONAL ANTIBODY - 21384742138474
      RABBIT ANTI-ACETYLCHOLINE POLYCLONAL ANTIBODY - 24467022446702
      RABBIT ANTI-ACETYLCHOLINE POLYCLONAL ANTIBODY2874326
      RABBIT ANTI-ACETYLCHOLINE POLYCLONAL ANTIBODY - 20427142042714

      References

      Reference overviewPub Med ID
      Neurotransmitter synthesis in poststroke cortical neurogenesis in adult rats.
      Gu, W; Gu, C; Jiang, W; Wester, P
      Stem cell research  4  148-54  2010

      Show Abstract
      20089468 20089468
      Immunocytochemical studies of the distribution of acetylcholine in the crayfish brain.
      Wang-Bennett, L T, et al.
      J. Comp. Neurol., 273: 330-43 (1988)  1988

      Show Abstract
      3209732 3209732
      One perfusion mixture for immunocytochemical detection of noradrenaline, dopamine, serotonin and acetylcholine in the same rat brain.
      McRae-Degueurce, A and Geffard, M
      Brain Res., 376: 217-9 (1986)  1986

      Show Abstract
      3719371 3719371
      Anti-acetylcholine antibodies and first immunocytochemical application in insect brain.
      Geffard, M, et al.
      Neurosci. Lett., 57: 1-6 (1985)  1985

      Show Abstract
      3897911 3897911