Microwave Antigen Retrieval Technique
From Shi, Shan-Rong et al, "Antigen retrieval technique:
A novel approach to immunohistochemistry on routinely processed
tissue sections,", in Gu, J, ed. ANALYTICAL
MORPHOLOGY: THEORY, LIBRARY AND PROTOCOLS, Eaton Publishing,
H2550* Laboratory MW Processor (Energy Beam Sciences)
- This MW processor with temperature readouts is available. Its
power output is regulated by a temperature feedback mechanism and
a timer, and it is capable of monitoring both temperature and heating
- Fix a Coplin jar on a thick capboard or plastic plate and set
it in the center portion of the turntable, filling the jar with
distilled water or tap water. Set the temperature probe into the
Coplin jar through a hole in the cap to measure the temperature.
- Set all test jars around the central probe jar as closely as
- Turn on the MW processor. Set the temperature and the time at
temperature as required.
- Turn on the turntable, making sure that the jars are not moved
by the probe and the table.
- Start the MW heating; the timer is automatically controlled.
- For heating at 100 degrees C, the heating time should be divided
into 5-minute cycles
- This MW processor is particularly useful for test battery (see
Test battery to develop an optimal protocol of AR for certain antibody
tested (Shi, S.-R., et al: Cell Vision 2:2-17, 1995)
- Ten slides numbered 1 to 10 are used for three pH values of AR
solution and three different heating conditions as follows:
A tenth slide stands as a non-AR control.
- Super-high temperature: Place slides in the microwave oven at
100 degrees C for 4 x 5 min....Kigh temperature: Microwave at 100
degrees C for 10 min as in the original protocol. Mid-high temperature:
Place in a temperature controlled MW oven...at 90 degrees C for
10 min. If possible, a H2550 MW processor may be used to maintain
the heating conditions accurately, as listed above.
- The use of an autostainer is recommended to perform IHC staining.
- Evaluate the intensity for all 10 slides in order to identify
the optimal protocol for the antibody under test.
- Additional studies, including different AR solutions...may be
used if necessary.
The of microwave energy as a means of pretreating formalin-fixed
tissue is a technique protected by U.S. Patent No. 5,244,787 assigned
to BioGenex Laboratories. Anyone seeking a license to practice this
technology should contact BioGenex directly.