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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| R | IHC | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MAB5300 |
| Brand Family | Chemicon® |
| Trade Name |
|
| Description | Anti-Dopamine Antibody, clone K56A |
| Product Information | |
|---|---|
| Format | Purified |
| Presentation | Purified immunoglobulin. Liquid containing 50% glycerol. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-Dopamine Antibody, clone K56A detects level of Dopamine & has been published & validated for use in IH. |
| Key Applications |
|
| Application Notes | Immunohistochemistry: 1:500-1:2,500 using free floating sections by the PAP technique on rat dopaminergic areas. Optimal working dilutions must be determined by end user. PROTOCOL for Dopamine 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 DOPAMINE 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, +4°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. 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. For research use only; not for use as a diagnostic. |
| Biological Information | |
|---|---|
| Immunogen | Dopamine-Gluteraldehyde-BSA. |
| Clone | K56A |
| Host | Mouse |
| Specificity | Dopamine. The cross-reactivities were determined using an ELISA test by competition experiments with the following compounds: Compound Cross-reactivity Dopamine-G-BSA 1 L-DOPA-G-BSA 1/10,000 Tyrosine-G-BSA 1/36,000 Tyramine-G-BSA 1/>50,000 Noradrenaline-G-BSA 1/>50,000 Octopamine-G-BSA 1/>50,000 Adrenaline-G-BSA 1/>50,000 Dopamine 1/>50,000 Abbreviations: (G) Glutaraldehyde (BSA) Bovine Serum Albumin |
| Isotype | IgG |
| Species Reactivity |
|
| Antibody Type | Monoclonal Antibody |
| Product Usage Statements | |
|---|---|
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Maintain at -20°C in undiluted aliquots for up to 6 months after date of receipt. |
| Packaging Information | |
|---|---|
| Material Size | 100 µL |