CD3 (CD3E) Antikörper

Antigen: CD3 (CD3E)

Synonyme: T3E, TCRE, FLJ18683

Klonalität: Monoklonal (F7238)

Wirt: Maus

Reaktivität: Human

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

Zertifikate: ISO 9001:2008, ISO 13485:2003

Anbieter: Dako

Produktnummer: ABIN370747

Produktbeschreibung

Weitere Bezeichnung: CD3

Immunogen: Purified CD3 epsilongammadelta/CD3ω (1).

Isotyp: IgG1, kappa chain  (Passende Sekundärantikörper)

Klon: F7238

Beschreibung: Synonyms for antigen T3, CD3 complex (2). The CD3 complex is composed of six polypeptides with usually four different transmembrane CD3 chains, gamma (gamma), delta (delta), epsilon (epsilon), and zeta (zeta). Three different dimers, gammaepsilon, deltaepsilon, and zetazeta, constitute the CD3 complex. The Mr of CD3epsilon is 20 000 (2). CD3 is first detectable in early thymocytes and its appearance probably represents one of the earliest signs of commitment to the T-cell lineage (3). In cortical thymocytes, the antigen is predominantly present as an intracytoplasmic constituent. It appears subsequently, at the medullar thymocyte stage, on the T-cell surface in close association with the T-cell receptor (TCR). Whereas the TCR forms the ligand-binding part of the TCR/CD3 complex, the function of the CD3 molecule is that of signalling, making CD3 a highly specific marker for T cells. No other cells are known to express the CD3 molecule, although two monoclonal antibodies raised against CD3 have been found to react with Purkinje cells in the cerebellum (4). The CD3 molecule is present in the great majority of T-cell neoplasms, although occasional tumours are encountered in which the antigen is lost as part of the neoplastic process (5). The CD3 molecule is also expressed in some cases of malignant histiocytosis (6) and Hodgkins lymphoma (7). CD3 epsilon, and not the whole CD3 molecule, has been detected in the cytoplasm of natural killer (NK) cells (8), and CD3epsilon may be a marker of nasal T-cell lymphomas which are thought to be of NK cell origin (9).

Spezifität: In Western blotting of FOX B cells transfected with CD3 epsilon cDNA (T116.4.3) or Jurkat E6.1 T cells, the antibody labels a 20 kDa band under reducing conditions, corresponding to CD3 epsilon. No labelling was observed in non-transfected FOX B cells (1). The antibody is most likely directed against the cytoplasmic region of the CD3 epsilon chain and works in immunocytochemistry as well as the DakoCytomation polyclonal Rabbit Anti- Human CD3, code No. A 0452 (1).

Anwendungen

Applikationshinweise: Paraffin sections: The antibody can be used for labelling paraffin-embedded tissue sections fixed in formalin, Duboscq-Brasil, Bouin's, or Bouin Holland (1). Pre-treatment of tissues with heat-induced epitope retrieval is required. For tissues fixed in formalin, optimal results are obtained with DakoCytomation Target Retrieval Solution, High pH, code No. S 3308, or 10 mmol/L Tris buffer, 1 mmol/L EDTA, pH 9.0. Less optimal results are obtained with DakoCytomation Target Retrieval Solution, code No. S 1700, or 10 mmol/L citrate buffer, pH 6.0 and pre-treatment of tissues with proteinase K. 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 frozen sections (1), and cell smears. Dilution: Monoclonal Mouse Anti-Human CD3, code No. M 7254, may be used at a dilution range of 1:25-1:50 when applied on formalin-fixed, paraffin-embedded sections of human tonsil and using 20 minutes heat-induced epitope retrieval in 10 mmol/L Tris buffer, 1 mmol/L EDTA, pH 9.0, 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. Automation: The antibody is well-suited for immunocytochemial staining using automated platforms (1), such as the DakoCytomation Autostainer. Cells labelled by the antibody display surface and/or cytoplasmic staining. Normal tissues: In thymus the antibody strongly labels lymphoid cells in the medulla and cortex. In tonsil and lymph node, interfollicular areas containing the T lymphocytes are strongly labelled, as also small reactive T cells in the B-cell follicles. With the exception of scattered T cells, all other normal human tissues studied were negative with the antibody (1). Abnormal tissues: The majority of T-cell lymphomas, 41/52 cases, were labelled by the antibody. Of non-Hodgkin's lymphomas, the antibody labelled 10/10 precursor T-lymphoblastic lymphomas/leukaemias, 11/11 peripheral T-cell lymphomas, 8/8 mycosis fungoides/Sezary syndrome, 2/2 angioimmunoblastic T-cell lymphomas (AILD), 1/1 intestinal T-cell lymphoma, 6/14 anaplastic large cell lymphomas (ALCL), 3/5 extranodal NK/T-cell lymphomas, nasal type, and 0/1 T-cell large cell granular lymphocytic leukaemia. In cases of Hodgkin's lymphoma, only 1/6 nodular sclerosis, and 1/1 unclassified showed focal cytoplasmic staining, whereas none of 5 cases of lymphocytic predominance, and 9 cases of mixed cellularity were labelled. None of 37 cases of different B-cell lymphomas were labelled (1).

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.

Forschungsgebiet: CD Antigene, Oberflächenrezeptoren der Immunzellen

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

Publikationen

Publikationen: Vela, Silva, Assad et al.: "Histopathological study of healing after allogenic mesenchymal stem cell delivery in myocardial infarction in dogs." in: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, Vol. 57, Issue 2, pp. 167-76, 2009 (PubMed).

Laghi, Bianchi, Miranda et al.: "CD3+ cells at the invasive margin of deeply invading (pT3-T4) colorectal cancer and risk of post-surgical metastasis: a longitudinal study." in: The lancet oncology, Vol. 10, Issue 9, pp. 877-84, 2009 (PubMed).

Helfer-Hungerbuehler, Cattori, Boretti et al.: "Dominance of highly divergent feline leukemia virus A progeny variants in a cat with recurrent viremia and fatal lymphoma." in: Retrovirology, Vol. 7, Issue 1, pp. 14, 2010 (PubMed).

Vezzali, Parodi, Marcato et al.: "Histopathologic classification of 171 cases of canine and feline non-Hodgkin lymphoma according to the WHO." in: Veterinary and comparative oncology, Vol. 8, Issue 1, pp. 38-49, 2010 (PubMed).

Lalor, Curbishley, Adams: "Identifying homing interactions in T-cell traffic in human disease." in: Methods in molecular biology (Clifton, N.J.), Vol. 616, pp. 231-52, 2010 (PubMed).