Immunohistochemical staining antibodies

Due to the relatively high-molecular-weight of the enzyme, conjugates formed with antibodies and P-gal tend to be much bulkier than those associated with AP or horseradish peroxidase. For this reason, antibody conjugates made with P-gal may have more difficulty penetrating tissue structures during immunohistochemical staining techniques than those made with the other enzymes. 

An additional control for the antibody specificity is the so-called absorption or preabsorption control, in which the primary antibody (prior to its use) is incubated for 1 h with a tenfold molar excess of the purified antigen. Absent or greatly diminished immunostaining should be obtained after application of this preabsorbed antibody. However, it is sometimes difficult to obtain the purified antigen therefore, it is rarely used routinely in immunohistochemical staining. Moreover, absorption of the antibody with the purified antigen does not always indicate that the antibody has bound to the same protein in the tissue (Burry 2000). 

Incubate in the primary antibody and complete immunohistochemical staining steps as desired. 

Light and electron microscopic studies were performed on the synovial membranes (E2) of patients with HIV associated arthropathy. An immunoperoxidase technique with the use of monoclonal antibodies against CD4, CD8, B, and DR lymphocytes and HIV p 24 antigen was also used. Mild to moderate nonspecific proliferative changes and increased vascularity of the subsynovial space were seen. Immunohistochemical staining revealed HIV p 24 positive staining cells of the synovial lining layer and the mononuclear cells of the subsynovial space, CD4, CD8 with predominance of CD8, B, and DR cells were also present. 

E. Ishikawa, M. Imagawa, S. Hashida, S. Yoshitake, Y. Hamaguchi, andT. Ueno, Enzyme-labeling of antibodies and their fragments for enzyme immunoassay and immunohistochemical staining, J. Immunoassay 4, 209-327 (1983). 

In neurohistochemistry, antibodies are used to provide extensive maps of the nervous system and to pinpoint antigens to specific locations and subpopulations of cells in the brain (Bloom et al., 1986 MacMillan and Cuello, 1986). In most laboratories, the use of immunohistochemical stains has become... 

Immunohistochemical stains use antibodies to identify specific constituents in tissue sections. In order to detect the site of reaction, the antibody is labeled with an enzyme that can be reacted with a suitable substrate to give a colored product. The alternative is to use a fluorescent label. The advantage of an enzyme label is that the nuclei can be counter stained thereby revealing the tissue architecture, and that the stain fades slowly, if at all, allowing the slides to be stored. 

Ericsson, 1990), the synthesis of fluorescently labeled DNA probes (L. M. Smith et al., 19 8 5), as a label in homogeneous immunoassay systems (Nithipatikom and McGown, 1987), to investigate specific interactions of proteins with cells surfaces (Hochman et al., 1988), and as an important fluorescent tag of antibodies in immunohistochemical staining techniques (Davidson and Hilchenbach, 1990). 

The ability to select the relative size of the antibody—enzyme complex is important, depending on the assay application. Low-molecular-weight conjugates may be more optimal for immunohistochemical staining or blotting techniques where penetration... 

Of the total number of commercial diagnostic assays utilizing antibody—enzyme conjugates, GO is employed in less than 1% of clinical tests. The enzyme remains, however, an important tool in many assays developed for research use. One particular advantage to the enzyme is that there is no endogenous GO activity in mammalian tissues, making it an excellent choice for immunohistochemical staining procedures. 

Feller, A. C., Parwaresch, M. R., Wacker, H.-H., Radzun, Fk-J., and Lennert, K. (1983) Combined immunohistochemical staining for surface IgD and T-lymphocyte subsets with monoclonal antibodies in human tonsils. Histochem. J. 15, 557—562. 

The ability of antibodies to bind molecules other than those molecules used as the immunogens is well known. Such a binding is termed cross-reactivity. Cross-reactivity is sometimes a source of confusion in the interpretation of immunohistochemically stained preparations because it cannot be detected by the usual controls for specificity. 

Immunohistochemical staining of tonsillitis, gastric adenocarcinomas, and breast carcinomas can be obtained using MIB-1 antibody in conjunction with EDTA-NAOH solution and a pressure cooker (Kim et ah, 1999b). EDTA solution is thought to be more effective than other buffers in unmasking the epitopes, presumably because it removes (chelates) tissue-bound calcium ions. 

Kammerer, U., Kapp, M., Gassel, A. M., Richter, T., Tank, C., Dietal, J., and Ruck, P. 2001. A new rapid immunohistochemical staining technique using the EnVision antibody complex. J. Histochem. Cytochem. 49 623-630. 

McCluggage, W. G., Maxwell, P., Patterson, A., and Sloan, J. M. 1997. Immunohistochemical staining of hepatocellular carcinoma with monoclonal antibody against inhibin. Histopathology 30 518-522. 

Mintze, K., Macon, N., Gould, K. E., and Sandusky, G. E., 1995. Optimization of proliferating cell nuclear antigen (PCNA). Immunohistochemical staining A comparison of methods using three commercial antibodies, various fixation times, and antigen retrieval solutions. J. Histotechnol. 78 25-30. 

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