Background: Flow cytometric analyses with monoclonal antibodies were so far mainly restricted to cell surface molecules. Intracellular structures such as cytoplasmic or nuclear enzymes, oncoproteins, cytokines, immunoglobulins etc. were largely excluded from such studies. Also excluded from flow cytometric studies were cytoplasmic localizations of well-established membrane molecules like CD3 and CD22, which, in their cytoplasmic form, are the most reliable lineage markers in undifferentiated leukemia. With the FIX&PERM® Kit flow cytometric analysis of intracellular antigens has become as easy as surface antigen studies. The only prerequisite is the availability of suitable antibody conjugates. Most of the available monoclonal antibody conjugates can be used with the FIX&PERM® Kit, some determinants are sensitive, however, to the fixation step involved. This and the optimal fixation time have to be tested for each reagent.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.
References: Recent application papers:Mestrum SGC, de Wit NCJ, Drent RJM, Hopman AHN, Ramaekers FCS, Leers MPG.Proliferative activity is disturbed in myeloproliferative neoplasms (MPN), myelodysplastic syndrome (MDS), and MDS/MPN diseases. Differences between MDS and MDS/MPN.Cytometry B Clin Cytom. 2021 May;100(3):322-330. doi: 10.1002/cyto.b.21946. Mestrum SGC, Cremers EMP, de Wit NCJ, Drent RJM, Ramaekers FCS, Hopman AHN, Leers MPG. Integration of the Ki-67 proliferation index into the Ogata score improves its diagnostic sensitivity for low-grade myelodysplastic syndromes. Leuk Res. 2022 Feb;113:106789. doi: 10.1016/j.leukres.2022.106789. Mestrum SGC, Cremers EMP, de Wit NCJ, Drent RJM, Ramaekers FCS, Hopman AHN, Leers MPG. Optimized gating strategy and supporting flow cytometry data for the determination of the Ki-67 proliferation index in the diagnosis of myelodysplastic syndrome. Data Brief. 2022 Feb 22;41:107976. doi: 10.1016/j.dib.2022.107976. Nies KPH, Kraaijvanger R, Lindelauf KHK, Drent RJMR, Rutten RMJ, Ramaekers FCS, Leers MPG. Determination of the proliferative fractions in differentiating hematopoietic cell lineages of normal bone marrow. Cytometry A. (2018) 93, 1097-1105 . doi: 10.1002/cyto.a.23564Further reading:- Gerna, G., Percivalle, E., Lilleri, D., Lozza, L., Fornara, C., Hahn, G., Baldanti, F. & Revello, M. G. (2005) J Gen Virol 86, 275-84.- Groeneveld, K., te Marvelde, J. G., van den Beemd, M. W., Hooijkaas, H. & van Dongen, J. J. (1996) Leukemia 10, 1383-9.- Haranaga, S., Yamaguchi, H., Friedman, H., Izumi, S., & Yamamoto, Y. (2001) Infect Immun 69, 7753-9.- Hegazy, A. N. & Klein, C. (2008) Leukemia 22, 2070-9.- Kappelmayer, J., Gratama, J. W., Karaszi, E., Menendez, P., Ciudad, J., Rivas, R. & Orfao, A. (2000) J Immunol Methods 242, 53-65.- Kline, M. P., Rajkumar, S. V., Timm, M. M., Kimlinger, T. K., Haug, J. L., Lust, J. A., Greipp, P. R. & Kumar, S. (2007) Leukemia 21, 1549-60- Knapp, W., Majdic, O. & Strobl, H. (1993) Recent Results Cancer Res 131, 31-40.- Knapp, W., Strobl, H. & Majdic, O. (1994) Cytometry 18, 187-98.- Knapp, W., Strobl, H., Scheinecker, C., Bello-Fernandez, C. & Majdic, O. (1995) Ann Hematol 70, 281-96.- Konikova, E., Glasova, M., Kusenda, J. & Babusikova, O. (1998) Neoplasma 45, 282-91.- Lanza, F., Latorraca, A., Moretti, S., Castagnari, B., Ferrari, L. & Castoldi, G. (1997) Cytometry 30, 134-44.- Millard, I., Degrave, E., Philippe, M. & Gala, J. L. (1998) Clin Chem 44, 2320-30.- Nakase, K., Sartor, M. & Bradstock (1998) Cytometry 34, 198-202.- Pascale, F., Contreras, V., Bonneau, M., Courbet, A., Chilmonczyk, S., Bevilacqua, C., Epardaud, M., Niborski, V., Riffault, S., Balazuc, A. M., Foulon, E., Guzylack-Piriou, L., Riteau, B., Hope, J., Bertho, N., Charley, B. & Schwartz-Cornil, I. (2008) J Immunol 180, 5963-72- Pickl, W. F., Majdic, O., Kohl, P., Stockl, J., Riedl, E., Scheinecker, C., Bello-Fernandez, C. & Knapp, W. (1996) J Immunol 157, 3850-9.- Riera-Sans, L., & Behrens, A. (2007) J Immunol 178, 5690-700- Roberts, J. L., Lengi, A., Brown, S. M., Chen, M., Zhou, Y. J., O'Shea, J. J. & Buckley, R. H. (2004) Blood 103, 2009-18- Sargent, R. L., Craig, F. E. & Swerdlow, S. H. (2009) Int J Clin Exp Pathol 2, 574-82- Scheinecker, C., Strobl, H., Fritsch, G., Csmarits, B., Krieger, O., Majdic, O. & Knapp, W. (1995) Blood 86, 4115-23.- Sedlmayr, P., Grosshaupt, B. & Muntean, W. (1996) Cytometry 23, 284-9.- Strobl, H. & Knapp, W. (2004) J Biol Regul Homeost Agents 18, 335-9.- Strobl, H., Scheinecker, C., Csmarits, B., Majdic, O. & Knapp, W. (1995) Br J Haematol 90, 774-82.- Strobl, H., Scheinecker, C., Riedl, E., Csmarits, B., Bello-Fernandez, C., Pickl, W. F., Majdic, O. & Knapp, W. (1998) J Immunol 161, 740-8.- Strobl, H., Takimoto, M., Majdic, O., Fritsch, G., Scheinecker, C., Hocker, P. & Knapp, W. (1993) Blood 82, 2069-78.- Wang, X., Chang, X., Facchinetti, V., Zhuang, Y. & Su, B. (2009) J Immunol 182, 3597-608
Caution: For professional users only.Reagent A of FIX&PERM® Cell Fixation and Permeabilization Kit contains fomaldehyde and is labelled: Harmful. Formaldehyde is toxic, allergenic and a suspected carcinogen. Never pipette by mouth and avoid contact with eyes, skin and clothing. Proper handling procedures are recommended. As a main rule, persons under 18 years of age are not allowed to work with this product. Users must be carefully instructed in the proper working procedure, the dangerous properties of the product and the necessary safety instructions. Please refer to the Material Safety Data Sheet (MSDS) for additional information.Dispose product remainders according to local regulations.
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