ActionsCite Favorites Display options Display options Format Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes) Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes) A murine monoclonal antibody specific for calf intestinal alkaline phosphatase has been prepared and used in an unlabeled antibody bridge technique for labeling monoclonal antibodies. This procedure--the alkaline phosphatase monoclonal anti-alkaline phosphatase (APAAP) method--gives excellent immunocytochemical labeling of tissue sections and cell smears, comparable in clarity and intensity to that achieved with immunoperoxidase labeling. If the enzyme label is developed with a naphthol salt as a coupling agent and Fast Red or hexazotized new fuchsin as a capture agent, a vivid red reaction product is obtained which is very easily detected by the human eye. For this reason the APAAP technique was found particularly suitable for labeling cell smears (for both cytoplasmic and surface-membrane antigens) and for detecting low numbers of antigen-bearing cells in a specimen (e.g., carcinoma cells in a malignant effusion). It was found possible to enhance the intensity of the APAAP labeling reaction substantially by repeating the second and third incubation steps (i.e., the unlabelled \"bridge\" antibody and APAAP complexes). The APAAP technique was superior to immunoperoxidase labeling for staining tissues rich in endogenous peroxidase, and could be used in conjunction with immunoperoxidase methods for double immunoenzymatic staining. The method was also applicable to the detection of antigenic molecules following their electrophoretic transfer from SDS-polyacrylamide gels to nitrocellulose sheets (\"immunoblotting\"). Falini B, Abdulaziz Z, Gerdes J, Canino S, Ciani C, Cordell JL, Knight PM, Stein H, Grignani F, Martelli MF, et al. Falini B, et al. J Immunol Methods. 1986 Nov 6;93(2):265-73. doi: 10.1016/0022-1759(86)90199-7. J Immunol Methods. 1986. PMID: 2430024 Schaumburg-Lever G. J Cutan Pathol. 1987 Feb;14(1):6-9. doi: 10.1111/j.1600-0560.1987.tb00120.x. J Cutan Pathol. 1987. PMID: 3549814 Pathol Immunopathol Res. 1988;7(3):169-86. doi: 10.1159/000157114. Pathol Immunopathol Res. 1988. PMID: 2461554 Adam M, Bekueretsion Y, Abubeker A, Tadesse F, Kwiecinska A, Howe R, Petros B, Jerkeman M, Gebremedhin A. Adam M, et al. JCO Glob Oncol. 2021 Feb;7:277-288. doi: 10.1200/GO.20.00391. JCO Glob Oncol. 2021. PMID: 33591838 Free PMC article. Hugenschmidt H, Labori KJ, Brunborg C, Verbeke CS, Seeberg LT, Bendigtsen Schirmer C, Renolen A, Borgen E, Naume B, Wiedswang G. Hugenschmidt H, et al. BMC Cancer. 2020 Nov 16;20(1):1107. doi: 10.1186/s12885-020-07510-z. BMC Cancer. 2020. PMID: 33198661 Free PMC article. Seidl M, et al. Histochem Cell Biol. 2020 Dec;154(6):663-669. doi: 10.1007/s00418-020-01906-5. Epub 2020 Aug 7. Histochem Cell Biol. 2020. PMID: 32767119 Free PMC article. Kerling A, Stoltenburg-Didinger G, Grams L, Tegtbur U, Horstmann H, Kück M, Mellerowicz H. Kerling A, et al. Orthop Res Rev. 2018 Aug 23;10:55-62. doi: 10.2147/ORR.S156449. eCollection 2018. Orthop Res Rev. 2018. PMID: 30774460 Free PMC article.