Antibodies immunohistochemistry

Avremeas S. Coupling of enzymes to proteins with glutaraldehyde use of the conjugates for the detection of antigens and antibodies. Immunohistochemistry 1971 6 394. 

Matcio L, Angelucci D, Grossi L, et al. Anti-Helicobacter pylori specific antibody immunohistochemistry improves the diagnostic accuracy of Helicobacter pylori in biopsy specimen from patients treated with triple therapy. Am J Gastroenterol. 1998 93 223-226. 

ADCC. Cetuximab is approved for treatment of metastatic colorectal cancer (CRC) and squamous cell carcinoma of the head and neck (SCCHN). Interestingly, an adverse event, acneiform rash seems to correlate with a better response to cetuximab, while there is no such correlation with expression levels of EGFR assessed by immunohistochemistry. Further side effects are rare infusion reactions and hypomagnesia. Two other anti-EGFR antibodies approved for clinical use are the fully human antibody panitumumab (Vectibix)... 

Mastocytosis is recognized in most patients because of the presence of characteristic cutaneous lesions [10]. A positive Darier s sign and/or histological examination of the skin using metachromatic stains, or by immunohistochemistry using antibodies to mast cell tryptase, helps confirm the diagnosis of cutaneous disease. 

Antibodies raised against the Ap peptide were subsequently used to identify more diffuse types of Ap-containing deposits in AD brain by immunohistochemistry. Diffuse Ap peptide deposits lack the... 

Hsi ED. A practical approach for evaluating new antibodies in the clinical immunohistochemistry laboratory. Arch. Pathol. Lab. Med. 2001 125 289-294. 

Shi SR, Imam SA, Young L, et al. Antigen retrieval immunohistochemistry under the influence of pH using monoclonal antibodies. J. Histochem. Cytochem. 1995 43 193-201. 

Ino H. Application of antigen retrieval by heating for double-label fluorescent immunohistochemistry with identical species-derived primary antibodies. J. Histochem. Cytochem. 2004 52 1219-1230. 

There are two types of specificity checks that may be warranted when choosing a specific peptide. The first is to demonstrate that the peptide is bound by the desired antibody and not by other, antigenically irrelevant antibodies. An example of this kind of specificity check is shown in Figure 7.3. A peptide that is immunoreactive with the 1D5 estrogen receptor (ER) mAb was covalently bound as a 1 pL spot to the center of each grid location. Various antibodies and controls were subsequently applied to the different grid locations. The bottom panel describes each of the antibodies that were applied to each grid location. The ability of the various antibodies to bind to the peptide was tested by immunohistochemistry. The presence of antibody bound to a... 

Montero C.The antigen-antibody reaction in immunohistochemistry./. Histochem. Cytochem. 2003 51 1-4. 

Boorsma, D.M., and Streefkerk, J.G. (1976b) Some aspects of the preparation, analysis, and use of peroxidase-antibody conjugates in immunohistochemistry. Protides Biol. Fluids, Proc. Colloq. 24, 795. 

Farr, A.C., and Nakane, P.K. (1981) Immunohistochemistry with enzyme labeled antibodies A brief review. J. Immunol. Meth. 47, 129-144. 

The use of protein A and protein G in immunohistochemistry is based on the same principle as that using secondary anti-IgG antibodies in the indirect two-step approach. In the first step, an antigen IgG complex is formed, which is then revealed in the second step by incubation with labeled protein A or protein G. 

The choice of fixation is a matter of experience, since the changes in the antigenicity of the component of interest resulting from the treatment with the fixative cannot be predicted. The epitope (amino acid sequence), against which the antibody was raised, may be hidden (masked), modified or completely lost as a result of an inadequate fixation (Shi et al. 2001). No ideal fixative has been found that can be used universally in immunohistochemistry. The choice of the adequate... 

It has also been proposed that new monoclonal antibodies be first examined by immunohistochemistry in several dilutions of twofold increments beyond those generally recommended by vendors, using 0.05 M Tris buffers of pH 6.0 and 8.6 without NaCl. Thereby, many monoclonal antibodies could be used in dilutions up to eightfold higher than those recommended by the vendor (Boenisch 1999). To the time of this writing, we have not yet had the opportunity to evaluate this apparently interesting approach. 

Boenisch T (1999) Diluent buffer ions and pH their influence on the performance of monoclonal antibodies in immunohistochemistry. Appl Immunohistochem Mol Morphol 7 300 306 Boenisch T (2005) Effect of heat induced antigen retrieval following inconsistent formalin fixation. Appl Immunohistochem Mol Morphol 13 283 286... 

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. 

Immunohistochemistry is a powerful method for identification of proteins in cells and tissues, but control procedures are necessary. The specificity of the results depends on two independent criteria the specificity of the antibody and of the method used (Burry 2000). The antibody specificity is best determined by immu-noblot and/or immunoprecipitation. The specificity of the method is best determined by both a negative control, replacing the primary antibody with the corresponding IgG (must be the same species as primary antibody at the same concentration), and a positive control, testing the primary antibody with tissues known to contain the antigen. 

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