Heat induced epitope retrieval

Despite some success with the above pretreatments, the development of wet heat-induced epitope retrieval (HIER) procedures has been the critical... 

The most commonly accepted point of view is that heat is responsible for unmasking the epitopes. In fact, Battifora (1996) has introduced the phrase heat-induced epitope retrieval (HIER). Heating at KXPC is a powerful treatment that can unmask hidden, buried, or crosslinked epitopes. Heat can be provided not only by a microwave oven, but also by an autoclave, a pressure cooker, steam, or a hot plate. A consensus on which method of heating is most effective in the retrieval of all types of epitopes is lacking. Therefore, some... 

The combined heat-induced epitope retrieval (HIER) and sonication-induced epitope retrieval (SIER) methods have been employed for retrieving cyclin Dl/bc 1-1 epitope in... 

Kim, Y.-S., Lee, S. J., Kim, I., Kim, G.-M., and Paik, S. R. 1999b. The use of alkaline EDTA solution improves heat-induced epitope retrieval for immunohistochemical localization of MIB-1 antigen. Acta Histochem. Cytochem. 32 287-294. 

Miller, R. T., and Estran, C. 1995. Heat-induced epitope retrieval with a pressure cooker. Appl. 

Shi, S.-R., Au, A., Soriano, R., Zhao, J.-X., and Sweet, W. L. 2000a. Non-biotin amplification (NBA) kit prevents nonspecific background staining of endogenous biotin induced by heat induced epitope retrieval (HIER) procedure. J. Histotechnol. 23 327-331. 

Bl. Bagdi, E., Krenacs, L., Krenacs, T., Miller, K., and Isaacson, P. G., Follicular dendritic cells in reactive and neoplastic lymphoid tissues A reevaluation of staining patterns of CD21, CD23, and CD35 antibodies in paraffin sections after wet heat-induced epitope retrieval. Appl. Immuno-histochem. Mol. Morphol. 9, 117—124 (2001). 

Sometimes AGEs have been localised using formalin-fixed paraffin-embedded tissue sections after heat-induced epitope retrieval, a process that could be responsible for the conversion of Amadori products to AGEs. Hayashi et al.401 therefore compared the reactivity of a monoclonal antibody to CML as an important AGE in... 

Dysbindin-1 proves to be a ubiquitous neuronal protein in the CNS under optimal immunohisto-chemical conditions. Those conditions include heat-induced epitope retrieval in EDTA (Pileri et al., 1997) and signal amplification using nickel sulfate (Cullen, 1994) or silver-gold (Teclemeriam-Mesbah et al., 1997). Virtually all neuronal populations appear to be immunoreactive for dysbindin-1 in the brain and the spinal cord (O Figure 2.2-13). As noted earlier, Iijima et al. (2009) reported that dysbindin-1 is found in... 

Figure 2. Coronin 1C in melanoma. A) Immunohistochemical staining of the indicated tissue samples for Coronin 1C. S m sections from melanoma tumors were stained with a new Coronin 1C specific mAb (Roadcap and Bear, unpublished reagent) for 30min at 37°C with a steam heat-induced epitope retrieval, using the streptavidin/biotin method with an alkaline phosphatase label and Permanent Red (Dako) as the chromogen and then stained with hematoxylin. B) qRT-PCR analysis of the indicated melanoma cell lines treated with the Erk inhibitor U0126 or vehicle control for 24 hours.Dotted line indicates lx threshold.

Riera JR, Astengo-Osuna C, Longmate JA, Battifora H. The immunohistochemical diagnostic panel for epithelial mesothelioma a reevaluation after heat-induced epitope retrieval. Am J Surg Pathol. 1997 21 1409-1419. 

Excludes neuroendocrine lung neoplasms. HIER, heat-induced epitope retrieval. 

Using formalin-hxed paraffin-embedded tissue sections, Hurlimann reported immunohistochemical desmin expression in 9 of 16 (56.3%) mesotheliomas (8 epithelial and 1 biphasic). Staining was described as focal or was found only in rare tumor cells. Cases showed positive reactivity more frequently with heat-induced epitope retrieval. Four of 16 (25%) mesotheliomas (3 epithelial and 1 biphasic) expressed neuron-specific eno-lase, 5 of 16 (31.3%) immunostained for chromogranin (4 epithelial, 1 biphasic) and 5 of 16 (31.3%) mesotheliomas (all epithelial) were positive for S-100 protein. Only rare tumor cells expressed these neuroepithelial markers. 

Key words Heat-induced epitope retrieval, Archived tissues, Restoring epitope conformation,... 

Despite some successes with the above pretreatments, the development of wet heat-induced epitope retrieval (HIER) procedures, which involves heating the fixed tissue sections in dilute metal-salt or buffer solutions at or above 100°C, for several minutes to 1/2 h, was the critical breakthrough in paraffin section immunohistochemistry (2, 7-9). Today, there are many variations of the original HIER technique. These differ primarily in the recommended buffer solutions and/or the source or mode of heating, but the basic formula of wet heat treatment over a fixed time period is similar. The most popular HIER technologies use microwave ovens, stainless steel or plastic pressure cookers, autoclaves, vegetable steamers or water-baths as the heat sources and low molarity buffers with acidic or alkaline pH (8,9,11-14). 

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