Antigen Retrieval

Enzymatic Method

  • Place dewaxed slides into ultrapure water at 37°C to warm them.

  • Make up enzyme solution.

    • Warm another 100ml ultrapure water to 37°C

    • Add 0.1g CaCl

    • Add 0.1g alpha-chymotrypsin (type II)

    • Adjust pH to 7.8

    • Warm to 37°C

  • Place warmed slides from ultrapure water into enzyme solution for 20 mins (no longer than 30min)

  • Remove slides and put into cold running tap water to stop the reaction.

  • Continue with immuno staining protocol.

Pressure Cooker Method

  • Dissolve 5.88g Tri-Sodium Citrate, and 44ml 0.2M HCL in 1956ml ultrapure water.

  • Adjust pH to 6 with 0.5% NaOH

  • Place solution in pressure cooker and apply heat until boiling, do not secure the lid yet.

  • Transfer the dewaxed slides to the pressure cooker and secure the lid.

  • Cook at full pressure for 3mins (5mins max).

  • Cool the slides.

  • Continue the labelling protocol

Microwave Method

  • Add 2.94g Tris-Sodium Citrate and 22ml 0.2M HCl to 978ml ultrapure water

  • Adjust to pH 6 with NaOH

  • Place de-waxed slides into microwavable dish and add solution.

  • Heat at full power for 15mins (30mins max).

  • Remove slides and allow to cool.

  • Continue with labelling procedure.