10X Citrate Buffer pH 8.0 Summary
| Specificity |
10x Citrate Buffer pH 8.0 for Heat Induced Epitope Recovery |
Applications/Dilutions
| Dilutions |
- Immunohistochemistry
- Immunohistochemistry-Paraffin
|
| Application Notes |
The antigen retrieval protocol is recommended for use in tissues that have been fixed in formalin only. Ensure that the fixed sections are adequately embedded in paraffin. Cut tissue sections to 4-5 microns.
Preparation of Working Solutions
1. The 10X concentrated format should be diluted tenfold with distilled or deionized water.
2. Mix one part of concentrated Antigen Retrieval Solution with nine parts of deionized or distilled water.
3. Shake the bottle vigorously to completely mix the components of the concentrate (the solution may separate into phases over time).
4. Store with cap tightly secured.
Protocol Recommendations
1. Deparaffinize and rehydrate tissue sections.
2. Place slides into 1X retrieval solution in a slide container (e.g. Coplin jar, Tissue-Tek, staining dish or metal slide canister).
3. Retrieve sections under pressure.
4. After take-off reagent jar containing slides from pressure cooker, allow the slides to cool for 20 minutes to reach room temperature.
5. Wash slides in deionized water and then with wash buffer. Proceed with immunostaining recommendations in the antibody datasheet.
6. Gently rinse by gradually adding DI water to the solution, then remove slides and rinse with DI water. |
Packaging, Storage & Formulations
| Storage |
Store at room temperature. |
| Buffer |
Dilute one part buffer with nine parts distilled water. |
| Preservative |
0.05% Sodium Azide |
Background
In order to perform immunostaining, the tissue specimens should be fixed in appropriate fixative. The purpose of such fixative is to conserve the tissue from autolysis, to preserve tissue structures and to immobilize antigens. However, this requires harsh treatment of the antigens. As a result, antigens undergo chemical alteration of their primary, secondary and tertiary structures. Because of changes in the protein containing epitopes or in neighboring proteins, antigenic sites may be masked. In past, enzymatic treatment with proteolytic enzymes i.e. pepsin, trypsin or pronase has been performed to regain the masked antigens. Shi et al. (1991) have reported that the treatment of the tissue section with heavy metal solution in a microwave can regain the masked antigens significantly. However, heavy metals in the solution increase the risk of exposure to lead for lab personnel. To solve this problem, we have developed a new antigen unmasking solution which is free from heavy metals. Use of this antigen unmasker can prevent the risk of unnecessary exposure for lab personnel and also resolve the handling and disposal problem.
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Support products are
guaranteed for 6 months from date of receipt.
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