CD65s antibody (FITC)

Antigen: CD65s

Clonality: Monoclonal (VIM2)

Host: Mouse

Reactivity: Human

Conjugate: FITC

Application: Flow Cytometry (FACS), Immunofluorescence (IF)

Catalog no.: ABIN132126

Additional Information

Format: Purified

Isotype: IgM

Clone: VIM2

Specificity: The epitope recognized by antibody VIM2 is expressed by virtually all myeloid cells including normal and malignant granulocytes and monocytes. In normal myelopoiesis VIM2 can first be detected after the late CFU-GM stage. In acute myeloid leukemias (AMLs) in vitro clonogenic progenitors seem to aberrantly express the VIM2 antigen. A variety of studies have demonstrated the usefulness and reliability of VIM2 as a marker molecule for the classification of acute leukemias. Recently, the signal transducing capacity of VIM2 bearing surface molecules has been demonstrated. Antibody VIM2 reacts with a carbohydrate structure expressed by myeloid cells. The epitope recognized was shown by Macher et al. to involve a defined sialofucooligosaccharide sequence. Similar results were obtained by Kniep et al.. Together with other sialylated and fucosylated polylactosamines the carbohydrate structure recognized by VIM2 may play a critical role on the adhesion of granulocytes and monocytes to endothelium and platelets during inflammation and clotting.

Application Details

Application Notes: The VIM2 antibody permits the identification and enumeration of normal and leukemic cell populations expressing the VIM2 antigen present in human biological samples (blood, bone marrow and others) using flow cytometry. Furthermore, VIM2 mAb is suitable for the elimination of myeloid cells from complex cell mixtures as well as for functional studies. (Lund-Johansen et al.) Results must be put within the context of other diagnostic tests as well as the clinical history of the patient by a certified professional before final interpretation. Analyses performed with this antibody should be paralleled by positive and negative controls. If unexpected results are obtained which cannot be attributed to differences in laboratory procedures, please contact us. Samples: Biological fluids (blood, bone marrow, and others) must be collected under sterile conditions. Anticoagulation with EDTA or heparin is recommended. The samples should be stored at room temperature until used. For optimal results, samples should be processed and analyzed within 24 hours. Samples with high numbers of non-viable cells might cause false results, such cases require determination of cell viability with e.g. propidium iodide. Sensitivity: The sensitivity of VIM2 mAb is determined by staining well-defined blood samples from representative donors with serial-fold mAb dilutions to obtain a titration curve that allows relating the mAb concentration to the percentage of stained cells and geometric MFI (mean fluorescence intensity). For this purpose, a mAb-concentration range is selected to include both the saturation point (i.e. the mAb dilution expected to bind all epitopes on the target cell) and the detection threshold (i.e. the mAb dilution expected to represent the least amount of mAb needed to detect an identical percentage of cells). In practice, 50ul of leukocytes containing 107cells/ml are stained with 20ul mAb of various dilutions to obtain a titration curve and to identify the saturation point and detection threshold. The final concentration of the product is then adjusted to be at least 3-fold above the detection threshold. In addition and to control lot-tolot variation, the given lot is compared and adjusted to fluorescence standards with defined intensity.

Purification: Chromatography

Buffer: PBS pH 7.2, 1% BSA, 0.05% NaN3

Storage: For stability reasons this monoclonal antibody solution contains sodium azide. These reagents should be stored at 2-8

Restrictions: For Research Use. CE-certified for In Vitro Diagnostics (IVD) Use in the European Union

Images

anti-CD65s antibody (FITC)

Publications

Publications: Lund-Johansen, Olweus, Horejsi et al.: "Activation of human phagocytes through carbohydrate antigens (CD15, sialyl-CD15, CDw17, and CDw65)." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 148, Issue 10, pp. 3221-9, 1992 (PubMed).

Macher, Beckstead: "Distribution of VIM-2 and SSEA-1 glycoconjugate epitopes among human leukocytes and leukemia cells." in: Leukemia research, Vol. 14, Issue 2, pp. 119-30, 1990 (PubMed).

Lowe, Kukowska-Latallo, Nair et al.: "Molecular cloning of a human fucosyltransferase gene that determines expression of the Lewis x and VIM-2 epitopes but not ELAM-1-dependent cell adhesion." in: The Journal of biological chemistry, Vol. 266, Issue 26, pp. 17467-77, 1991 (PubMed).

Springer, Lasky: "Cell adhesion. Sticky sugars for selectins." in: Nature, Vol. 349, Issue 6306, pp. 196-7, 1991 (PubMed).

Schwarzinger, Valent, Koeller et al.: "Prognostic significance of surface marker expression on blasts of patients with de novo acute myeloblastic leukemia." in: Journal of clinical oncology : official journal of the American Society of Clinical Oncology, Vol. 8, Issue 3, pp. 423-30, 1990 (PubMed).

Uemura, Macher, DeGregorio et al.: "Glycosphingolipid carriers of carbohydrate antigens of human myeloid cells recognized by monoclonal antibodies." in: Biochimica et biophysica acta, Vol. 846, Issue 1, pp. 26-36, 1985 (PubMed).

Koeller, Haas, Ludwig et al.: "Phenotypic and genotypic heterogeneity in infant acute leukemia. II. Acute nonlymphoblastic leukemia." in: Leukemia : official journal of the Leukemia Society of America, Leukemia Research Fund, U.K, Vol. 3, Issue 10, pp. 708-14, 1989 (PubMed).

Macher, Buehler, Scudder et al.: "A novel carbohydrate, differentiation antigen on fucogangliosides of human myeloid cells recognized by monoclonal antibody VIM-2." in: The Journal of biological chemistry, Vol. 263, Issue 21, pp. 10186-91, 1988 (PubMed).

Delwel, Bot, Knapp et al.: "Vim-2, candidate monoclonal antibody for purging autologous marrow grafts in acute myeloblastic leukaemia." in: Bone marrow transplantation, Vol. 2, Issue 2, pp. 149-54, 1988 (PubMed).

Lutz, Krieger, Bettelheim et al.: "[Clinical value of various classifications in adults with AML]" in: Onkologie, Vol. 9, Issue 2, pp. 67-71, 1986 (PubMed).

Bettelheim, Panzer, Majdic et al.: "Unexpected absence of a myeloid surface antigen (3-fucosyl-N-acetyllactosamine) in promyelocytic leukemia." in: Leukemia research, Vol. 9, Issue 11, pp. 1323-7, 1986 (PubMed).

Peschel, Konwalinka, Geissler et al.: "Studies on differentiation of committed hemopoietic progenitor cells with monoclonal antibodies directed against myeloid differentiation antigens." in: Experimental hematology, Vol. 13, Issue 11, pp. 1211-6, 1986 (PubMed).

Majdic, Bettelheim, Stockinger et al.: "M2, a novel myelomonocytic cell surface antigen and its distribution on leukemic cells." in: International journal of cancer. Journal international du cancer, Vol. 33, Issue 5, pp. 617-23, 1984 (PubMed).

Knapp, Strobl, Majdic: "Flow cytometric analysis of cell-surface and intracellular antigens in leukemia diagnosis." in: Cytometry, Vol. 18, Issue 4, pp. 187-98, 1995 (PubMed).

Stoeckl, Majdic, Rosenkranz et al.: "Monoclonal antibodies to the carbohydrate structure Lewis(x) stimulate the adhesive activity of leukocyte integrin CD11b/CD18 (CR3, Mac-1, alpha m beta 2) on human granulocytes." in: Journal of leukocyte biology, Vol. 53, Issue 5, pp. 541-9, 1993 (PubMed).

Lund-Johansen, Olweus, Symington et al.: "Activation of human monocytes and granulocytes by monoclonal antibodies to glycosylphosphatidylinositol-anchored antigens." in: European journal of immunology, Vol. 23, Issue 11, pp. 2782-91, 1993 (PubMed).

Fink, Koeller, Mayer et al.: "Prognostic significance of myeloid-associated antigen expression on blast cells in children with acute lymphoblastic leukemia. The Austrian Pediatric Oncology Group." in: Medical and pediatric oncology, Vol. 21, Issue 5, pp. 340-6, 1993 (PubMed).

Clarke, Watkins: "Alpha1,3-L-fucosyltransferase expression in developing human myeloid cells. Antigenic, enzymatic, and mRNA analyses." in: The Journal of biological chemistry, Vol. 271, Issue 17, pp. 10317-28, 1996 (PubMed).

Knibbs, Craig, Natsuka et al.: "The fucosyltransferase FucT-VII regulates E-selectin ligand synthesis in human T cells." in: The Journal of cell biology, Vol. 133, Issue 4, pp. 911-20, 1996 (PubMed).

Kniep, Peter-Katalinic, Muething et al.: "The CDw65 monoclonal antibodies VIM-8 and VIM-11 bind to the neutral glycolipid V3FucnLc8Cer." in: Journal of biochemistry, Vol. 119, Issue 3, pp. 456-62, 1996 (PubMed).

Wagers, Stoolman, Kannagi et al.: "Expression of leukocyte fucosyltransferases regulates binding to E-selectin: relationship to previously implicated carbohydrate epitopes." in: Journal of immunology (Baltimore, Md. : 1950), Vol. 159, Issue 4, pp. 1917-29, 1997 (PubMed).

Noguchi, Sato, Sugimori et al.: "A minor E-selectin ligand, CD65, is critical for extravascular infiltration of acute myeloid leukemia cells." in: Leukemia research, Vol. 25, Issue 10, pp. 847-53, 2001 (PubMed).

Watkins, Clarke: "The genetic regulation of fucosylated and sialylated antigens on developing myeloid cells." in: Advances in experimental medicine and biology, Vol. 491, pp. 231-65, 2003 (PubMed).

Brown, Clarke, Ledbetter et al.: "Elevated expression of adenosine A1 receptor in bronchial biopsy specimens from asthmatic subjects." in: The European respiratory journal : official journal of the European Society for Clinical Respiratory Physiology, Vol. 31, Issue 2, pp. 311-9, 2008 (PubMed).