To one ml of buffer (reagent B), add 2-4 drops of concentrated enzyme (reagent E) in a test tube, mix well. This ready-to-use reagent is good for 2-3 hours.
1. Deparafinize FFPE tissue section is carried out as outlined in your protocol. Hydrate tissue with PBS.
2. Remove buffer, add 2-4 drops of ready-to use enzyme solution to cover the tissue section.
3. Incubate at 37ºC for 5-10 minutes or 10-15 minutes at room temperature, (the time required for optimum digestion of FFPE tissue section will vary with the extent of fixation, generally approx. 5-15 minutes are sufficient at the above temperatures. In some cases it may require more time).
4. Discard trypsin reagent, wash 3-5 times with buffer.
5. The tissue sections are ready for further IHC protocol.
Store at 2-8ºC, DO NOT FREEZE.
For laboratory research use only. Not for any clinical, therapeutic, or diagnostic use in humans or animals. Not for animal or human consumption.
Purchasers shall not, and agree not to enable third parties to, analyze, copy, reverse engineer or otherwise attempt to determine the structure or sequence of the product.
Trypsin is used for proteolytic digestion of formalin-fixed paraffin-embedded (FFPE) tissue sections prior to application of antibodies. In IHC most commonly used fixative like formalin; mask tissue antigens (cellular, membrane and nuclear) by cross-linking process, this results in poor or no staining in IHC. Trypsin digestion improves immunoreactivity of some antigens in FFPE tissue sections. For cytokeratin clone AE3 and AE1/AE3, this enzyme works much better than boiling the tissue with citrate buffer.