Pepsin Solution For Enzyme-Induced Epitope Retrieval
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Specificity and Comments: Formaldehyde fixation impairs or totally destroys the immunoreactivity of many antigens and epitopes. The negative effect of formaldehyde fixation can be reversed successfully with enzymatic digestion for some markers while not for others. Digestion of deparaffinized tissue sections with the enzyme pepsin has been found to improve the immunoreactivity of many antigens in formaldehyde fixed tissues.
Supplied As: Pepsin solution (pH2.0) is supplied as a single-component in a ready-to-use format for immnuohistochemical staining of formalin-fixed, paraffin-embedded tissues.
Step-by-Step Instructions: - Place five micron thick tissue sections on glass slides coated with poly L-lysine or APTES.
- Deparaffinize and re-hydrate sections as usual.
- Block endogenous peroxidase as usual.
- Wash sections in wash-buffer for 2x5 minutes.
- Cover sections with the pepsin solution (usually 0.2ml per section) and place the slides at 37oC for 10 minutes in a humidified chamber.
- Wash sections in wash buffer for 2x5 minutes
- Block non-specific sites with normal serum as usual.
- Place optimally diluted primary antibody on the sections (incubation time and temperature for a given set of experimental conditions should be determined by the investigator).
- Wash sections in buffer for 2x5 minutes.
- Rest of the procedure is same as routinely performed in your laboratory.
Suggested Test Size: It is recommended that at least 0.2ml of pepsin solution should be used for each tissue section.
Storage and Stability: Store vial at 4°C. When stored at 2-8°C, this pepsin solution is stable for 12 months.
Limitations and Warranty: The pepsin solution is available for research use only and is not approved for use in humans or in clinical diagnosis. There are no warranties, expressed or implied, which extend beyond this description. NeoMarkers is not liable for property damage, personal injury, or economic loss caused by this product.
Custom Service: Please contact us if you require this product in a special format.
Material Safety Data: This product is not licensed or approved for administration to humans or to animals other than the experimental animals. Standard Laboratory Practices should be followed when handling this material. The chemical, physical, and toxicological properties of this material have not been thoroughly investigated. Appropriate measures should be taken to avoid skin and eye contact, inhalation, and ingestion.
