Trypsin Solution For Enzyme-Induced epitope Retrieval
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Description: 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 trypsin has been found to improve the immunoreactivity of some antigens in formaldehyde fixed tissues.
Supplied As: Trypsin solution is supplied as a two-component system. Before use, mix 3-drops of solution-A with 1-drop of solution-B for digestion of formalin-fixed, paraffin-embedded tissue sections.
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.
- Before use, mix 3-drops of Trypsin Solution-A with 1-drop of Trypsin Solution-B
- Cover sections with the pre-mixed trypsin solution (usually 0.2ml per section) and place the slides at 37C 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 trypsin solution should be used for each tissue section.
Storage and Stability: Store vial at 4°C. When stored at 2-8°C, this trypsin solution is stable for 12 months.
