Immunoperoxidase

Taylor CR, Mason DY. The immunohistological detection of intracellular immunoglobulin in formalin-paraffin sections from multiple myeloma and related conditions using the immunoperoxidase technique. Clin. Exp. Immunol. 1974 18 417 29. 

DeLellis RA, Sternberger LA, Mann RB, et al. Immunoperoxidase technics in diagnostic pathology. Report of a workshop sponsored by the National Cancer Institute. Am. J. Clin. Pathol. 1979 71 483-488. 

Human embryo lung fibroblasts infected with a reference laboratory strain of herpes simplex virus (HS V) type 2 were used to detect antibody to HSV type 2 in serum samples. After treatment of cells with serial dilutions of sera, HRP-labeled immunoglobulins to human IgG (class G immunoglobulins) were added and detected with CL substrate [36], In both cases a sharp detection of the specific antibodies was achieved with chemiluminescent assays, which proved more sensitive than the colorimetric immunoperoxidase assays. 

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. 

Giorno R. 1984. A comparison of two immunoperoxidase staining methods based on the avidin-biotin interaction. Diagn Immunol 2 161-166. 

Hsu SM, Raine L, Fanger H. 1981. Use of avidin-biotin-per-oxidase complex (ABC) in immunoperoxidase techniques a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 29 577-580. 

Figure 25. A-D Immunofluorescence staining of vmculrn in vascular smooth muscle cells on day 3 after seeding on polymeric surfaces (medium supplemented with 10% fetal bovine serum). A poly(DL-lactic acid), PDLLA B block copolymer of poly(DL-lactic acid) and poly (ethylene oxide) (PEO), PDLLA-6-PEO C, E PDLLA-6-PEO with 5% GRGDSG-PEO-6-PDLLA D, F PDLLA-6-PEO with 20% GRGDSG-PEO-6-PDLLA. E, F Immunoperoxidase staining of bromodeoxyuridine (arrows) incorporated into DNA newly synthesized in vascular smooth muscle cells cultured for 3 days in serum-free medium on PDLLA-Z)-PEO with 5% (E) or 20% (F) GRGDSG-PEO-6-PDLLA. Cells counterstained with light green. Bar=100 pm [41].

Login, G., Schnitt, S., and Dvorak, A. (1987) Rapid microwave fixation of human tissues for light microscopic immunoperoxidase identification of diagnostically useful antigens. Lab. Invest. 57, 585-591. 

Milde, P., Merke, J., Ritz, E., Haussler, M., and Rauterberg, E. (1989) Immuno-histochemical detection of 1,25-dihydroxyvitamin D3 receptors and estrogen receptors by monoclonal antibodies comparison of four immunoperoxidase methods. J. Histochem. Cytochem. 37,1609-1617. 

Hsu, S. M., Raine, L., and Fanger, H. (1981) The use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase technique a comparison between ABC and unlabeled antibody PAP procedures. J. Histochem. Cytochem. 29, 577-580. 

Discussed herein are the general requirements of the immunohistochemistry laboratory for compliance with CLIA-88. In addition, a troubleshooting guide for immunoperoxidase procedures is included. 

When analyzing an immunostained specimen, deposits of the colored chromogen indicate the presence of the antigen and represent specific positive staining. The pattern of staining in the cells can be cytoplasmic, nuclear, membranous, or surface focal or diffuse. This section addresses some of the most common problems encountered in immunoperoxidase procedures and the appropriate solutions to correct them (see Chapters 23-25). (See Note 4 for additional troubleshooting resources.)... 

Improper fixation and processing Proper antigen fixation is the cornerstone of immunoperoxidase techniques, for without it, results will be poor. Some markers may be destroyed by certain fixatives overfixation may mask certain antigens (see Chapter 8) (4). Review manufacturer s package inserts for appropriate fixation techniques. Paraffin-embedded tissue should never be exposed to temperatures >60°C as this can destroy some antigens. 

The benefit of the immunoperoxidase technique is best exhibited by the ability to detect neoplastic B cells. By demonstrating the presence of k or X (but not both) in lymphoid or plasma cells, one can establish a diagnosis of lymphoma or plasmacytoma. Unfortunately, in routinely processed tissue, only immunoglobulins in the cytoplasm and not on the cell surface membrane can be detected. 

Kamino, H. and Tam, S. T. (1991) Immunoperoxidase technique modified by counterstain with azure B as a diagnostic aid in evaluation heavily pigmented melanocytic neoplasms./. Cutaneous Pathol. 18, 436 39. 

G3. Glynn, P, Gilbert, H., Newcombe, J., and Cuzner, M. L., Rapid analysis of immunoglobulin isoelectric focusing patterns with cellulose nitrate sheets and immunoperoxidase staining. J. Immunol. Methods 51, 251-257 (1982). 

This chapter describes a simple immunoperoxidase method for detection of PCNA in tissue sections. A flow cytometric method for demonstrating PCNA in single cells is given in Chapter 36. 

Accurate quantitation of antigens using immunohistochemistry depends upon a linear relationship between the amount of antigen and the intensity of immunoperoxidase-DAB reaction product as well as the percentage of stained cells. Variations in staining intensity will reflect the amount of antigen only if optimal preparatory procedures are used for example, oversaturation of the chromogen reaction may result in invalid quantitation. Therefore, optimal concentration of DAB should be determined by trials with DAB... 

Recently, an automatic color video image analysis system was developed to quantify antigen expression (androgen receptor) (Kim et al.,T999a). This system provides a linear relationship between the antigen content and mean optical density of the immunoperoxidase-substrate reaction product. Titration of antibody, concentration, and reaction duration of the substrate can be optimized with this system. The imaging hardware consists of a Zeiss microscope, a three-chip charge-coupled-device camera, a camera control board, and a Pentium-based personal computer. 

A cocktail (1 1) of two monoclonal anticyclin Dl/bc 1-1 antibodies were used P2D11F11 (diluted 1 40) and 5D4 (diluted 1 100) these two antibodies can be obtained from Vecta Laboratories, Inc., Burlingame, CA, and Immunotech, Westbrook, ME, respectively. The avidin-biotin immunoperoxidase detection system employing DAB is used as the chromogen (Ventana Biotek). After counterstaining in dilute Mayer s hematoxylin, the sections are dehydrated and mounted in Permount. 

Simultaneous detection of multiple antigens provides a spatial relationship between the antigens of interest and saves time, effort, and tissue specimens. In the standard immunoperoxidase technique, horseradish peroxidase is used to oxidize the colorless chromogen DAB into a brown end-product in the presence of hydrogen peroxide. When nickel chloride is included in the reaction mixture, the final reaction product is black. By... 

Boon, M. E., Kok, L. P., Moorlag, H. E., and Suurmeijer, A. J. 1989. Accelerated immunogold silver immunoperoxidase staining of paraffin sections with the use of microwave irradiation. Am, J. Clin. Pathol. 92 137-143. 

---