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品种反应性 主要应用 宿主 格式 抗体类型 R IHC M Purified Monoclonal Antibody 描述 产品目录编号 MAB5304 品牌系列 Chemicon®
商名 - Chemicon
描述 Anti-Glutamate Antibody 产品信息 格式 Purified 演示 Liquid in PBS containing 10mM sodium azide. 应用 应用 Anti-Glutamate Antibody detects level of Glutamate & has been published & validated for use in IH. 主要应用 - Immunohistochemistry
应用说明 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.生物信息 免疫原品种 Glutamate-Glutaraldehyde-BSA. 宿主 Mouse 特异性 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.同种型 IgM 品种反应性 Rat 抗体类型 Monoclonal Antibody 产品使用声明 使用声明 - 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.
储存和货运信息 存储条件 Maintain at 2-8°C in undiluted aliquots for up to 6 months after date. 包装信息 数量 100 µL
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