IHC antigen retrieval reagent (citrate, pH 7.0) is used to recover antigens masked by fixation in cross linking fixatives such as formalin.
Product Details
Formulation: | Liquid. 10X citrate buffer, pH 7.0. |
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Applications: | IHC
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Shipping: | Ambient Temperature |
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Long Term Storage: | Ambient |
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Technical Info/Product Notes: | In order to perform immunostaining, tissue specimens should be preserved in an appropriate fixative. Fixation stops tissue autolysis, preserves tissue structures, and immobilizes antigens. However, antigens undergo chemical alteration of their primary, secondary and tertiary structures during fixation. Antigenic sites may be masked due to changes induced in the epitope or neighboring proteins. Enzymatic treatment with proteolytic enzymes (i.e. pepsin, trypsin or pronase) has been performed to expose the masked antigens. Shi et al. (1991) have reported that treating tissue sections with a heavy metal solution in a microwave oven can cover masked antigens significantly. However, heavy metals in the solution increase the risk of toxic exposure to lab personnel. To reduce the risk, we have developed an antigen unmasking solution which is free of heavy metals. The reagent is provided in a convenient 10X concentrate. Use of this antigen recovery buffer avoids unnecessary heavy metal exposure to lab personal and other handling and disposal issues.
Interpretation of the results will be the sole responsibility of the user. |
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Protocol: | Preparation of reagent: Dilute one part buffer with nine parts distilled water.
Procedure:
1. Deparaffinize and rehydrate tissue sections.
2. Fill a coplin jar with sufficient 1x Citrate Buffer to cover the tissue sections on the slides.
3. Place coplin jar in steamer or water bath.
4. Heat steamer or water bath containing coplin jar to 95-100°C.
5. Place deparaffinized slides (1-3 slides/jar) in the coplin jar and incubate for 20-40 minutes (optimal incubation time should be determined by the end user).
6. Remove coplin jar from the water bath and allow the slides to cool for 20 minutes to reach room temperature.
7. Wash slides in deionized water and then with wash buffer. Proceed with immunostaining. |
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Regulatory Status: | RUO - Research Use Only |
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Product Literature References
Effect of T3 Spinal Contusion Injury on Upper Urinary Tract Function: J.H. Gumble, et al.; Neurotrauma Rep.
3, 190 (2022),
Abstract;
General Literature References
Antigen retrieval in formalin-fixed, paraffin-embedded tissues: an enhancement method for immunohistochemical staining based on microwave oven heating of tissue sections: S.R. Shi, et al.; J. Histochem. Cytochem.
39, 741 (1991),
Abstract;
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