Trypsin Reagent

Trypsin is an antigen retriever solution 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.

1. Deparafinize FFPE tissue section is carried out as outlined in your protocol. Hydrate tissue with PBS or ImmunoBioScience Universal Immuno buffer. Add endogenous peroxidase block as usual. Wash tissue section 3-5 times with buffer. 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.

Trypsin is an antigen retriever solution 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.