Clone: VIM2
Label: FITC
Background: 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. 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.
Source: VIM2 was produced after immunization of mice with an extract of THP-1, a monocytic cell line derived from a patient with acute monocytic leukemia.
Specificity: 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. 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 fluorescenceintensity). 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 10^7cells/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 three-fold above the detection threshold. In addition and to control lot-to-lot variation, the given lot is compared and adjusted to fluorescence standards with defined intensity.
Formulation: PBS pH 7.2, 1% BSA, 0.05% NaN3
References: 1. O. Majdic et al., Int J Cancer 33, 617 (1984). 2. P. Bettelheim et al., Leuk Res 9, 1323 (1985). 3. C. Peschel et al., Exp Hematol 13, 1211 (1985). 4. K. Uemura et al., Biochim Biophys Acta 846, 26 (1985). 5. D. Lutz et al., Onkologie 9, 67 (1986). 6. R. Delwel, F. Bot, W. Knapp, B. Lowenberg, Bone Marrow Transplant 2, 149 (1987). 7. B. A. Macher, J. Buehler, P. Scudder, W. Knapp, T. Feizi, J Biol Chem 263, 10186 (1988). 8. U. Koller et al., Leukemia 3, 708 (1989)9. B. A. Macher, J. H. Beckstead, Leuk Res 14, 119 (1990). 10. I. Schwarzinger et al., J Clin Oncol 8, 423 (1990). 11. J. B. Lowe et al., J Biol Chem 266, 17467 (1991). 12. T. A. Springer, L. A. Lasky, Nature 349, 196 (1991). 13. F. Lund-Johansen et al., J Immunol 148, 3221 (1992). 14. F. M. Fink et al., Med Pediatr Oncol 21, 340 (1993). 15. F. Lund-Johansen et al., Eur J Immunol 23, 2782 (1993). 16. J. Stockl et al., J Leukoc Biol 53, 541 (1993). 17. W. Knapp, H. Strobl, O. Majdic, Cytometry 18, 187 (1994). 18. G. M. Brown, T. N. Huckerby, B. L. Abram, I. A. Nieduszynski, Biochem J 319 ( Pt 1), 137 (1996). 19. J. L. Clarke, W. Watkins, J Biol Chem 271, 10317 (1996). 20. R. N. Knibbs et al., J Cell Biol 133, 911 (1996). 21. B. Kniep et al., J Biochem (Tokyo) 119, 456 (1996). 22. A. J. Wagers, L. M. Stoolman, R. Kannagi, R. Craig, G. S. Kansas, J Immunol 159, 1917 (1997). 23. M. Noguchi, N. Sato, H. Sugimori, K. Mori, K. Oshimi, Leuk Res 25, 847 (2001). 24. W. M. Watkins, J. L. Clarke, Adv Exp Med Biol 491, 231 (2001).
Caution: For professional users only. This reagent contains sodium azide. To avoid the development of hazardous conditions, reagents containing azide should be diluted in running water prior to be discarded. Similar to the work with other biological products, proper handling procedures are recommended.
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