MAB5304

Anti-Glutamate Antibody

Chemicon^®, from mouse

• UNSPSC-Code: 12352203
• eCl@ss: 32160702
• NACRES: NA.41

Eigenschaften

• Biologische Quelle: mouse
• Antikörperform: purified antibody
• Antikörper-Produkttyp: primary antibodies
• Klon: monoclonal
• Speziesreaktivität: rat
• Hersteller/Markenname: Chemicon^®
• Methode(n): immunohistochemistry: suitable
• Isotyp: IgM
• Versandbedingung: dry ice
• Posttranslationale Modifikation Target: unmodified

Beschreibung

Spezifität

Glutamate

The cross-reactivities were determined using an ELISA test by competition experiments with the following compounds:

Compound Cross-reactivity

Glutamate-G-BSA 1

Aspartate-G-BSA 1/100,000

GABA-G-BSA 1/100,000

Glutamate 1/100,000

Abbreviations:

(G) Glutaraldehyde

(BSA) Bovine Serum Albumin

NOTE: Antibody reactivity REQUIRES glutaraldehyde fixation thus some glutaraldehyde (0.5%-2.0%) needs to be included in the tissue fixation procedure inorder for the proper reactivity.

Immunogen

Glutamate-Glutaraldehyde-BSA.

Anwendung

Research Category
Neurowissenschaft

Research Sub Category
Neurotransmitter & Rezeptoren

Neuroinflammation & Schmerz

Anti-Glutamate Antibody detects level of Glutamate & has been published & validated for use in IH.

Immunohistochemistry: 1:2,500-1:10,000 using free floating sections by the PAP technique on rat hippocampus.

Optimal working dilutions must be determined by end user.

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

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

Solution A: 0.1M cacodylate, 10g/L sodium metabisulfite, pH 6.2.

Solution B: 0.1M cacodylate, 10g/L sodium metabisulfite, 3-5% glutaraldehyde, pH 7.5.

2. RAT PERFUSION - The rat is anaesthetized with sodium pentobarbital or Nembutal and perfused intracardially through the aorta using a pump with Solution A (30 mL): 150-300 mL/min, Solution B (500 mL): 150-300 mL/min.

3. POST FIXATION: 15 to 30 minutes in Solution B, then 4 soft washes in 0.05M Tris with 8.5 g/L sodium metabisulfite, pH 7.5 (Solution C) .

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

5. REDUCTION STEP: Sections are reduced with Solution C containing 0.1M sodium borohydride for 10 minutes. The sections are washed 4 times in solution C without sodium borohydride.

6. APPLICATION OF GLUTAMATE ANTIBODY: Use a final dilution of 1:2,500-1:10,000 in Solution C containing 0.1% Triton X100 and 2% non-specific serum. Incubate 12 sections per 2 mL diluted antibody overnight, +2-8°C. Then wash the sections three times for 10 minutes each in Solution C. (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.

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

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

9. 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. Mount sections on chrome-alum coated slides. Dry overnight at 37°C. Rehydrate sections using conventional histological procedures. Coverslip using rapid mounting media.

For research use only; not for use as a diagnostic.

Physikalische Form

Format: Purified

Liquid in PBS containing 10mM sodium azide.

Lagerung und Haltbarkeit

Maintain at 2-8°C in undiluted aliquots for up to 6 months after date.

Rechtliche Hinweise

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Haftungsausschluss

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.

Sicherheitshinweise

• Lagerklassenschlüssel: 12 - Non Combustible Liquids
• WGK: WGK 2
• Flammpunkt (°F): Not applicable
• Flammpunkt (°C): Not applicable