Immunohistochemistry methods

Correspondingly, two principal immunohistochemistry methods are employed direct and indirect. The direct method is a straightforward one-step process that creates a direct reaction between the antigen and the labeled antibody. The indirect method requires the use of two antibodies a primary unlabeled antibody and a secondary labeled antibody. 

First-line chemotherapy for the treatment of metastatic breast cancer is now largely dependent on the patient s HER-2 antigen status. The HER-2 (human epidermal growth factor receptor 2) protein is overexpressed on the surface of breast cancer cells in approximately 15-25% of patients. The measurement of this protein can be performed by immunohistochemistry methods, where the degree of protein expression is denoted by negative, 1+, 2+ or 3+. As Mrs CR s has HER-2,3+ disease, this denotes strong protein overexpression. 

DNLM 1. Immunohistochemistry—methods. 2. Diagnostic Techniques and Procedures. 3. Neoplasms—diagnosis. QW 504.5 D536 2009] RB46.6.D33 2009... 

Renshaw S (2007) Immunohistochemistry Methods Express Series. Oxford, UK Scion Publishing Ltd. 

Immunohistochemistry is a powerful and very useful tool in diagnostic pathology consequently, the staining protocol must be optimized to ensure that the results are reproducible, reliable and accurate. A standard immunohistochemistry method includes the following steps ... 

Other than expression within blood vessels within the malignant tissue, no expression was observed in melanomas and lymphomas using this three-step immunohistochemistry method. Paraffin sections showed a marked loss of expression, as did pre-incubation of frozen sections with methanol, indicating that the NeuSGc epitopes are likely to be present mostly on glycolipids. 

Taylor CR, Levenson RM. Quantification of immunohistochemistry—issues concerning methods, utility and semiquantitative assessment II. Histopathology 2006 49 411 124. 

Shi S-R, Liu C, Young L, et al. Development of an optimal antigen retrieval protocol for immunohistochemistry of retinoblastoma protein (pRB) in formalin fixed, paraffin sections based on comparison of different methods. Biotech. Histochem. 2007 82 301-309. 

Wilson E, Jackson S, Cruwys S, et al. An evaluation of the immunohistochemistry benefits of boric acid antigen retrieval on rat decalcified joint tissues. J. Immunol. Methods 2007 322 137-142. 

Frost AR, Sparks D, Grizzle WE. Methods of antigen recovery vary in their usefulness in unmasking specific antigens in immunohistochemistry. Appl. Immunohistochem. Mol. Morphol. 2000 8 236-243. 

Pileri SA, Roncador G, Ceccarelli C, et al. Antigen retrieval techniques in immunohistochemistry comparison of different methods. I. Pathol. 1997 183 116-123. 

Zeheb R. Automating immunohistochemistry. In Immunohistochemical Staining Methods, 4th edition, ed. ME Key, pp. 103-106. Carpinteria, CA Dako, 2006. 

Prior to beginning of IHC on formalin paraffin tissues more than 30 years ago, fresh cell smears or frozen tissue sections were used for immunofluorescence studies on tissues sections, and in a more limited way for immunperoxi-dase studies, now generally known as immunohistochemistry, prior to adaptation of the method to FFPE in 1974. The traditional viewpoint that... 

When fresh or frozen tissue is used for proteomic analyses, the results cannot be related directly to the clinical course of diseases in a timely manner. Instead, researchers frequently reduce the number of interesting proteins to a manageable number and then attempt to use immunohistochemistry to understand the implications of proteomic changes in archival formalin-fixed, paraffin-embedded (FFPE) tissue for which the clinical course has been established.3 Unfortunately, immunohistochemistry is a semiquantitative pro-teomic method, and the choice of interesting proteins must occur without advance knowledge of the clinical course of the disease or the response to therapy. If routinely fixed and embedded archival tissues could be used for standard proteomic methods such as 2-D gel electrophoresis and mass spectrometry (MS), these powerful techniques could be used to both qualitatively and quantitatively analyze large numbers of tissues for which the clinical course has been established. However, analysis of archival FFPE tissues by... 

Hyatt MA (2002) Microscopy, Immunohistochemistry, and Antigen Retrieval Methods For Light and Electron Microscopy. Plenum, New York... 

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