Mouse anti CD65s

Mouse anti CD65s

• 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.
• CAS Number: 9007-83-4
• Host: Mouse
• Species Reactivity: Human
• Isotype: IgM
• Clone: VIM2
• Type: Primary Antibodies
• Applications: Direct IF, IF, Flow Cytometry
• Assay Principle: Staining Procedure Direct Immunofluorescence (Staining Procedure) Nordic-MUbio fluorochrome labeled antibodies are designed for use with either whole blood or isolated mononuclear cell (MNC) preparations. Proposed staining procedure for whole blood in short: - For each sample add 50 µL of EDTA anti-coagulated blood to a 3-5 mL tube - Add 20 µL of the appropriate Nordic-MUbio monoclonal antibody conjµgate - Incubate the tube for 15 minutes at 4°C or at room temperature in the dark - Add 100 µL NM-LYSE (Cat.No. GAS-003) to each tube and incubate for 10 minutes at room temperature - Add 3-4 mL of destilled water and vortex, incubate for 5-10 minutes at room temperature - Centrifµge tube for 5 minutes at 300 g - Aspirate supernatant and resuspend pellet in 0.3 mL of sheath fluid - Analyze immediately or store samples at 2-8° C in the dark and analyze within 24 hours For “No-Wash” protocol please refer to www.nordicmubio.com Proposed staining procedure for MNC in short: - Carefully add 20 µL antibody conjµgate and 50-100 µL MNC to the bottom of a tube - Vortex at low speed for 1-2 seconds - Incubate for 15-30 minutes at 2-8°C or at room temperature - Centrifµge tubes for 5 minutes at 300 g - Remove supernatant, resuspend cells in 2-5 mL of phosphate buffered saline (PBS) and centrifµge cells again for 5 minutes at 300 g - Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 mL 1 % formaldehyde and store them at 2-8°C in the dark. Analyze fixed cells within 24 hours Indirect Immunofluorescence (Staining Procedure) - Mix 20 µL Nordic-MUbio purified antibody with 50 µL whole blood or MNC suspension - Incubate for 15 minutes at 2-8°C - Wash cells with phosphate buffered saline (PBS) - Add to cell pellet 20 µL of affinity purified, fluorochrome labeled F(ab’)2 anti mouse Ig antibodies - Incubate for 15 minutes at 2-8°C - Wash cells with phosphate buffered saline (PBS) or proceed as described for direct staining
• Form: PBS pH 7,2, 1% BSA, 0,05% NaN3
• Precautions: 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.
• References & Citations: 1. O. Majdic et al., Int J Cancer 33, 617 (1984). P. Bettelheim et al., Leuk Res 9, 1323 (1985). _x000D_ 2. C. Peschel et al., Exp Hematol 13, 1211 (1985). _x000D_ 3. K. Uemura et al., Biochim Biophys Acta 846, 26 (1985). _x000D_ 4. D. Lutz et al., Onkologie 9, 67 (1986). _x000D_ 5. R. Delwel, F. Bot, W. Knapp, B. Lowenberg, Bone Marrow Transplant 2, 149 (1987). _x000D_ 6. B. A. Macher, J. Buehler, P. Scudder, W. Knapp, T. Feizi, J Biol Chem 263, 10186 (1988). _x000D_ 7. U. Koller et al., Leukemia 3, 708 (1989)_x000D_ 8. B. A. Macher, J. H. Beckstead, Leuk Res 14, 119 (1990). _x000D_ 9. I. Schwarzinger et al., J Clin Oncol 8, 423 (1990). _x000D_ 10. J. B. Lowe et al., J Biol Chem 266, 17467 (1991). _x000D_ 11. T. A. Springer, L. A. Lasky, Nature 349, 196 (1991). _x000D_ 12. F. Lund-Johansen et al., J Immunol 148, 3221 (1992). _x000D_ 13. F. M. Fink et al., Med Pediatr Oncol 21, 340 (1993). _x000D_ 14. F. Lund-Johansen et al., Eur J Immunol 23, 2782 (1993). _x000D_ 15. J. Stockl et al., J Leukoc Biol 53, 541 (1993). _x000D_ 16. W. Knapp, H. Strobl, O. Majdic, Cytometry 18, 187 (1994). _x000D_ 17. G. M. Brown, T. N. Huckerby, B. L. Abram, I. A. Nieduszynski, Biochem J 319 ( Pt 1), 137 (1996). _x000D_ 18. J. L. Clarke, W. Watkins, J Biol Chem 271, 10317 (1996). _x000D_ 19. R. N. Knibbs et al., J Cell Biol 133, 911 (1996). _x000D_ 20. B. Kniep et al., J Biochem (Tokyo) 119, 456 (1996). _x000D_ 21. A. J. Wagers, L. M. Stoolman, R. Kannagi, R. Craig, G. S. Kansas, J Immunol 159, 1917 (1997). _x000D_ 22. M. Noguchi, N. Sato, H. Sugimori, K. Mori, K. Oshimi, Leuk Res 25, 847 (2001). _x000D_ 23. W. M. Watkins, J. L. Clarke, Adv Exp Med Biol 491, 231 (2001).
• Storage Conditions: Nordic-MUbio monoclonal antibody reagents contain optimal concentrations of affinity-purified antibody; For stability reasons this monoclonal antibody solution contains sodium azide. These reagents should be stored at 2-8°C (DO NOT FREEZE!) and protected from prolonged exposure to light. If a slight precipitation occurs upon storage, this should be removed by centrifugation; It will not affect the performance or the concentration of the product Stability of the reagent: Please refer to the expiry date printed onto the vial. The use of the reagent after the expiration date is not recommended.