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Electron Microscopy Sciences

Technical Data Sheets

Trypsin Reagent

EMS Catalog #64142-01, 64142-02

Introduction

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.

Storage

2-8°C, DO NOT FREEZE

Intended Use

Use as antigen retriever for some antibodies in IHC. Please refer to primary antibody protocol for IHC.

Reagent

This enzyme is supplied as concentrated form along with buffer.

Mixing of reagent

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.

The unused solution can be discarded according to city, county, state, province or country’s regulations.

Procedures

  1. Deparafinize FFPE tissue section is carried out as outlined in your protocol.
  2. Hydrate tissue with PBS or ImmunoBioScience Universal Immuno buffer
  3. Add endogenous peroxidase block as usual.
  4. Wash tissue section 3-5 times with buffer.
  5. Remove buffer, add 2-4 drops of ready-to use enzyme solution to cover the tissue section.
  6. Incubate at 37°C 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).
  7. Discard trypsin reagent, wash 3-5 times with buffer.
  8. The tissue sections are ready for further IHC protocol.

Limitation: Over-digestion may detach tissue sections from slides or may result in the loss of tissue morphology. It may be necessary to use + charge, sialanized or polylysine coated slides.

WARNING

This product contains 0.05% Proclin 300 as a preservative, appropriate care should be taken in handling. Avoid skin and eye contact with all laboratory products. Use appropriate laboratory gear, laboratory coat, gloves and safety glasses. Do not ingest any laboratory products. This product is not approved for administration in human or animals.

For research use only; not for use in diagnostic procedures. FOR IN VITRO LABORATORY USE ONLY

Country of origin: USA

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Trypsin Reagent Kit