Antigen Retrieval Techniques for use with Formalin-Fixed Paraffin-Embedded Sections
In many cases the fixation and processing steps involved in the preparation of tissue results in loss of antigen immunoreactivity. Often this can be reversed by using appropriate antigen retrieval techniques such as microwave antigen retrieval or proteolytic digestion.
Standard protocols for some of these techniques are outlined below. It should be noted that in many cases, incubation times will vary depending upon the particular processing method used, the antigen to be retrieved and/or the antibody being used.
This method provides a general procedure for use with the majority of Bio-Rad reagents. In some cases specific recommendations are provided on product datasheets, and these methods should always be used in conjunction with product and batch specific information provided with each vial. Please note that a certain level of technical skill and immunological knowledge is required for the successful design and implementation of these techniques - these are guidelines only and may need to be adjusted for particular applications.
These techniques can be used with the following protocols:
Heat-mediated antigen retrieval may be used in many situations to enable staining of certain antigens in paraffin-embedded tissue sections, or sometimes to improve results when staining is weak or inconsistent.
A number of buffers may be used for this purpose. The precise choice is usually recommended on product specific datasheets, or determined experimentally. For your convenience, Bio-Rad offers a range of pre-prepared antigen retrieval buffers at different pH levels guaranteeing consistency of performance and reliability. Alternatively, buffers may be prepared using the two recipes below. Please contact Bio-Rad’s Technical Services Department for more details.
Please note that the actual heating times may vary depending on the antibody and the fixation processes being used.