Epithelial Antigen Antikörper

Antigen: Epithelial Antigen

Klonalität: Monoklonal (Ber-EP4)

Wirt: Maus

Reaktivität: Human

Applikation: Immunhistochemie (Paraffinschnitte) (IHC (p))

Zertifikate: ISO 9001:2008, ISO 13485:2003

Anbieter: Dako

Produktnummer: ABIN370599

Produktbeschreibung

Immunogen: MCF-7 cells (human breast carcinoma cell line) (4).

Isotyp: IgG1, kappa chain

Klon: Ber-EP4

Beschreibung: Epithelial antigen is a cell surface glycoprotein of unknown function (4). This epithelium-specific antigen is broadly distributed in epithelial cells, and displays a highly conserved expression in carcinomas (4, 5). As exceptions to the general expression in normal epithelia, adult hepatocytes, in contrast to fetal hepatocytes, parietal cells in gastric glands, and apical cells in squamous epithelia are negative. Epithelial antigen is rarely present in mesotheliomas (1, 4). It has been reported that epithelial antigen plays an important role as tumour-cell marker in lymph nodes from patients with oesophageal carcinoma otherwise classified as node-negative (2). Epithelial antigen has also been suggested as a discriminator between basal cell and basosquamous carcinomas, and squamous cell carcinoma of the skin (3).

Spezifität: Analysis of immunocomplexes between the antibody and lysate of 125 I surface-labelled MCF-7 cells in SDS-PAGE under reducing conditions shows that the antibody labels two polypeptides of 34 kDa and 39 kDa, respectively, corresponding to epithelial antigen. Under non-reducing conditions, the polypeptides appear as 39 kDa and 41 kDa, while deglycosylation reduces the size to 31 kDa, and 36 kDa. In immunoprecipitation experiments, the Ber-EP4 antibody blocks the reaction of the HEA125 antibody with MCF-7 cell lysate and vice versa, showing that the two antibodies react with the same antigen. The two antibodies also produce identical staining results in cells and tissues (4). Of 37 cell lines tested, the antibody homogeneously labels all (10/10) carcinoma cell lines, whereas all non-epithelial cell lines (26/27) are negative except for the erythromyeloid cell line K562 (4).

Anwendungen

Applikationshinweise: Paraffin sections: The antibody can be used for labelling paraffin-embedded tissue sections fixed in formalin. Pre-treatment of tissues with heat-induced epitope retrieval is required. Optimal results are obtained with DakoCytomation Target Retrieval Solution, code No. S 1700. Less optimal results are obtained with 10 mmol/L citrate buffer, pH 6.0. However, DakoCytomation Target Retrieval Solution, High pH, code No. S 3308, or 10 mmol/L Tris buffer, 1 mmol/L EDTA, pH 9.0, and were found inefficient. Pre-treatment of tissues with proteinase K was found less optimal. The tissue sections should not dry out during the treatment or during the following immunocytochemical staining procedure. Frozen sections and cell preparations: The antibody can be used for labelling acetone-fixed, frozen sections and cell preparations (4). Dilution: Monoclonal Mouse Anti-Human Epithelial Antigen, code No. M 0804, may be used at a dilution range of 1:200-1:400 when applied on formalin-fixed, paraffin-embedded sections of human kidney and using 20 minutes heat-induced epitope retrieval in DakoCytomation Target Retrieval Solution, code No. S 1700, and 30 minutes incubation at room temperature with the primary antibody. Optimal conditions may vary depending on specimen and preparation method, and should be determined by each individual laboratory. The recommended negative control is DakoCytomation Mouse IgG1, code No. X 0931, diluted to the same mouse IgG concentration as the primary antibody. Unless the stability of the diluted antibody and negative control has been established in the actual staining procedure, it is recommended to dilute these reagents immediately before use, or dilute in DakoCytomation Antibody Diluent, code No. S 0809. Positive and negative controls should be run simultaneously with patient specimen. Visualization: DAKO LSAB/HRP kit, code No. K 0679, and DAKO EnVision/HRP kits, code Nos. K 4004 and K 4006, are recommended. For frozen sections and cell preparations, the DakoCytomation APAAP kit, code No. K 0670, is a good alternative if endogenous peroxidase staining is a concern. Follow the procedure enclosed with the selected visualization kit. Product-specific limitations Cells labelled by the antibody display membrane and cytoplasmic staining. The membrane staining is preferentially basolateral (4). In formalin-fixed, paraffin-embedded carcinomas arising from tissues with no or smaller amounts of epithelial antigen, such as hepatocellular and lung carcinomas, the antibody does not perform satisfactorily compared to labelling in frozen sections of the same tissues (4). Owing to the relative lability of the epitope in formalin-fixed, paraffin-embedded tissue sections, negative results should be interpreted with caution (4).

Buffer: Monoclonal mouse antibody provided in liquid form as cell culture supernatant dialysed against 0.05 mol/L Tris/HCl, pH 7.2, and containing 15 mmol/L NaN 3 .

Lagerung: Store at 2-8 °C. Precautions: 1. For professional users. 2. This product contains sodium azide (NaN 3 ), a chemical highly toxic in pure form. At product concentrations, though not classified as hazardous, sodium azide may react with lead and copper plumbing to form highly explosive build-ups of metal azides. Upon disposal, flush with large volumes of water to prevent metal azide build-up in plumbing. 3. As with any product derived from biological sources, proper handling procedures should be used.

Beschränkungen: Für Forschungszwecke. Als CE-zertifizierter Antikörper in der Europäischen Union für In Vitro Diagnostik (IVD) zugelassen.

Publikationen

Publikationen: Cras, Martin, Gheuens: "Gamma-enolase and glial fibrillary acidic protein in nervous system tumors. An immunohistochemical study using specific monoclonal antibodies." in: Acta neuropathologica, Vol. 75, Issue 4, pp. 377-84, 1988 (PubMed).

Soler-Federsppiel, Cras, Gheuens et al.: "Human gamma gamma-enolase: two-site immunoradiometric assay with a single monoclonal antibody." in: Journal of neurochemistry, Vol. 48, Issue 1, pp. 22-8, 1987 (PubMed).

Shimizu, Suzuki, Kato: "Characterization of alpha alpha, beta beta, gamma gamma and alpha gamma human enolase isozymes, and preparation of hybrid enolases (alpha gamma, beta gamma and alpha beta) from homodimeric forms." in: Biochimica et biophysica acta, Vol. 748, Issue 2, pp. 278-84, 1983 (PubMed).

Anstey, Cerio, Ramnarain et al.: "Desmoplastic malignant melanoma. An immunocytochemical study of 25 cases." in: The American Journal of dermatopathology, Vol. 16, Issue 1, pp. 14-22, 1994 (PubMed).

Song, Yang, Wei et al.: "[Allogeneic stem cell transplantation for 75 cases of acute myeloid leukemia in complete remission: outcome and prognostic analysis]" in: Zhongguo shi yan xue ye xue za zhi / Zhongguo bing li sheng li xue hui = Journal of experimental hematology / Chinese Association of Pathophysiology, Vol. 18, Issue 1, pp. 161-6, 2010 (PubMed).