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FIX&PERM(r) Cell Fixation and Permeabilization Kit

Research Use Only
GAS-002
Nordic-MUbio
Product group Technique Specific Products
Price on request
8 x 5 ml
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More than 130 suppliers
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Overview

  • Supplier
    Nordic-MUbio
  • Product Name
    FIX&PERM Cell Fixation and Permeabilization Kit
  • Delivery Days Customer
    7
  • Application Supplier Note
    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. All biological samples have to be handled with caution. Always consider them as potentially infective. Use appropriate precautions such as gloves, lab-coat, etc. Fixation, permeabilization and staining procedure: For each sample to be analyzed add 50 microl of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube Add 100 microl of Reagent A (Fixation Medium, stored and used at room temperature) Incubate for 15 minutes at room temperature Add 5ml phosphate buffered saline and centrifuge cells for 5 minutes at 300 g Remove supernatant and add to cell pellet 100 microl Reagent B (Permeabilization Medium) and 20 microl of the appropriate Nordic-MUbio monoclonal antibody conjugate Vortex at low speed for 1-2 seconds Incubate for 15 minutes at room temperature Wash cells with phosphate buffered saline as described above Remove supernatant and resuspend cells in sheath fluid for immediate analysis or resuspend cells in 0.5 ml 1.0 % formaldehyde and store them at 2-8°C in the dark. Analyze fixed cells within 24 hours. Comments: Special cases (diluted bone marrow samples, other samples containing low soluble protein) might benefit from replenishment with plasma components before the FIX&PERM® treatment in order to create a milieu, which more closely resembles the stuation in anti-coagulated blood. For that pupose addition of IgG preparations (e.g. Beriglobulin P, ZLB Behring, final concentration 10mg/ml) and human serum albumin (e.g. human albumin Behring 20% - infusion solution, final concentration 40mg/ml) is recommended.
  • Applications
    Flow Cytometry
  • Applications Supplier
    Flow Cytometry
  • Category Supplier
    Buffers and reagents
  • Certification
    Research Use Only
  • Storage Instruction
    RT
  • UNSPSC
    41116124

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.23564 Further 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 -Fan J, Tang X, Wang Q, Zhang Z, Wu S, Li W, Liu S, Yao G, Chen H and Sun L, Mesenchymal stem cells alleviate experimental autoimmune cholangitis through immunosuppression and cytoprotective function mediated by galectin-9, Stem Cell Research & Therapy 2018, https://doi.org/10.1186/s13287-018-0979-x Bingyu Shi, Jingjing Qi, Genhong Yao, Ruihai Feng, Zhuoya Zhang, Dandan Wang, Chen Chen, Xiaojun Tang, Liwei Lu, Wanjun Chen and Lingyun Sun Mesenchymal stem cell transplantation ameliorates Sjögren’s syndrome via suppressing IL-12 production by dendritic cells. Stem Cell Research & Therapy, 2018 https://doi.org/10.1186/s13287-018-1023 Rossella Zanin, Silvia Pegoraro, Gloria Ros, Yari Ciani, Silvano Piazza, Fleur Bossi, Roberta Bulla, Cristina Zennaro, Federica Tonon, Dejan Lazarevic, Elia Stupka, Riccardo Sgarra & Guidalberto Manfioletti HMGA1 promotes breast cancer angiogenesis supporting the stability, nuclear localization and transcriptional activity of FOXM1 Journal of Experimental & Clinical Cancer Researchvolume 38, Article number: 313 (2019) https://doi.org/10.1186/s13046-019-1307-8 Manabu Itoh, Yosuke Mukae, Takahiro Kitsuka, Kenichi Arai, Anna Nakamura, Kazuyoshi Uchihashi, Shuji Toda, Kumika Matsubayashi, Jun-ichi Oyama, Koichi Node, Daisuke Kami, Satoshi Gojo, Shigeki Morita, Takahiro Nishida, Koichi Nakayama & Eiji Kobayashi Development of an immunodeficient pig model allowing long-term accommodation of artificial human vascular tubes. Nature Communicationsvolume 10, Article number: 2244 (2019) https://doi.org/10.1038/s41467-019-10107-1