FIX&PERM Solution A (Fix) (CE) Reagent
FIX&PERM Solution A (Fix) (CE)
Applications
Flow Cytometry, Functional Assay
Synonyms
FIX&PERM Solution A (Fix) (CE); FIX&PERM Solution A (Fix) (CE) reagent
Specificity
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.
The quality of each FIX&PERM Lot is determined by fixation and permeabilization of well defined blood samples from representative donors and subsequent comparison of forward and side scatter characteristics of obtained leukocytes.
All biological samples have to be handled with caution. Always consider them as potentially infective. Use appropriate precautions such as gloves, lab-coat, etc.
The quality of each FIX&PERM Lot is determined by fixation and permeabilization of well defined blood samples from representative donors and subsequent comparison of forward and side scatter characteristics of obtained leukocytes.
Form/Format
1 x 100ml vial of Fixation Medium (Reagent A)
FIX&PERM Reagents are designed for use with all commercially available flow cytometers. Alignment and compensation should be performed according to manufacturer´s instructions.
FIX&PERM Reagents are designed for use with all commercially available flow cytometers. Alignment and compensation should be performed according to manufacturer´s instructions.
Applicable Applications for FIX&PERM Solution A (Fix) (CE) reagent
Flow Cytometry (FC/FACS)
Application Notes
1. For each sample to be analyzed add 50 ul of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube
2. Add 100 ul of Reagent A (Fixation Medium, stored and used at room temperature)
3. Incubate for 15 minutes at room temperature
4. Add 5ml phosphate buffered saline and centrifuge cells for 5 minutes at 300g
5. Remove supernatant and add to cell pellet 100 ul Reagent B (Permeabilization Medium) and 20 ul of the appropriate monoclonal antibody conjugate
6. Vortex at low speed for 1-2 seconds
7. Incubate for 15 minutes at room temperature
8. Wash cells with phosphate buffered saline as described above
9. 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 degree 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.
2. Add 100 ul of Reagent A (Fixation Medium, stored and used at room temperature)
3. Incubate for 15 minutes at room temperature
4. Add 5ml phosphate buffered saline and centrifuge cells for 5 minutes at 300g
5. Remove supernatant and add to cell pellet 100 ul Reagent B (Permeabilization Medium) and 20 ul of the appropriate monoclonal antibody conjugate
6. Vortex at low speed for 1-2 seconds
7. Incubate for 15 minutes at room temperature
8. Wash cells with phosphate buffered saline as described above
9. 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 degree 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.
CE Mark
CE
Preparation and Storage
Reagents should be stored and used at room temperature. Do not freeze. 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. If reagents are stored under any conditions other than those specified, the conditions must be verified by the user. Do not use reagents if a precipitate should form or discoloration occurs. If unexpected results are obtained which cannot be attributed to differences in laboratory procedures, please contact us.
Related Product Information for FIX&PERM Solution A (Fix) (CE) reagent
This FIX&PERM Cell Permeabilization Kit contains 2 reagents: Fixation Medium (Reagent A) and Permeabilization Medium (Reagent B). It is intended for first fixing cells in suspension with Reagent A and then permeabilizing the cell membranes with Reagent B. This procedure gives antibodies access to intracellular structures and leaves the morphological scatter characteristics of cells intact. Specific formulations reduce background staining and allow simultaneous addition of permeabilization medium and fluorochrome labeled antibodies.
FIX&PERM is suitable for the analysis of normal and malignant leukocyte populations derived from various human biological samples (blood, bone marrow and others) using flow cytometry. Results must be put within the context of other tests as well as the clinical history of the patient by a certified professional before final interpretation.
FIX&PERM is suitable for the analysis of normal and malignant leukocyte populations derived from various human biological samples (blood, bone marrow and others) using flow cytometry. Results must be put within the context of other tests as well as the clinical history of the patient by a certified professional before final interpretation.
References
Wang, X., Chang, X., Facchinetti, V., Zhuang, Y. & Su, B. (2009) J Immunol 182, 3597-608
Sargent, R. L., Craig, F. E. & Swerdlow, S. H. (2009) Int J Clin Exp Pathol 2, 574-82
Hegazy, A. N. & Klein, C. (2008) Leukemia 22, 2070-9
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
Riera-Sans, L., & Behrens, A. (2007) J Immunol 178, 5690-700
Gerna, G., Percivalle, E., Lilleri, D., Lozza, L., Fornara, C., Hahn, G., Baldanti, F. & Revello, M. G. (2005) J Gen Virol 86, 275-84.
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
Strobl, H. & Knapp, W. (2004) J Biol Regul Homeost Agents 18, 335-9.
Haranaga, S., Yamaguchi, H., Friedman, H., Izumi, S., & Yamamoto, Y. (2001) Infect Immun 69, 7753-9
Kappelmayer, J., Gratama, J. W., Karaszi, E., Menendez, P., Ciudad, J., Rivas, R. & Orfao, A. (2000) J Immunol Methods 242, 53-65.
Millard, I., Degrave, E., Philippe, M. & Gala, J. L. (1998) Clin Chem 44, 2320-30.
Strobl, H., Scheinecker, C., Riedl, E., Csmarits, B., Bello-Fernandez, C., Pickl, W. F., Majdic, O. & Knapp, W. (1998) J Immunol 161, 740-8.
Konikova, E., Glasova, M., Kusenda, J. & Babusikova, O. (1998) Neoplasma 45, 282-91.
Millard, I., Degrave, E., Philippe, M. & Gala, J. L. (1998) Clin Chem 44, 2320-30.
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.
Sedlmayr, P., Grosshaupt, B. & Muntean, W. (1996) Cytometry 23, 284-9.
Groeneveld, K., te Marvelde, J. G., van den Beemd, M. W., Hooijkaas, H. & van Dongen, J. J. (1996) Leukemia 10, 1383-9.
Knapp, W., Strobl, H., Scheinecker, C., Bello-Fernandez, C. & Majdic, O. (1995) Ann Hematol 70, 281-96.
Scheinecker, C., Strobl, H., Fritsch, G., Csmarits, B., Krieger, O., Majdic, O. & Knapp, W. (1995) Blood 86, 4115-23.
Strobl, H., Scheinecker, C., Csmarits, B., Majdic, O. & Knapp, W. (1995) Br J Haematol 90, 774-82.
Knapp, W., Strobl, H. & Majdic, O. (1994) Cytometry 18, 187-98.
Knapp, W., Majdic, O. & Strobl, H. (1993) Recent Results Cancer Res 131, 31-40.
Strobl, H., Takimoto, M., Majdic, O., Fritsch, G., Scheinecker, C., Hocker, P. & Knapp, W. (1993) Blood 82, 2069-78.
Sargent, R. L., Craig, F. E. & Swerdlow, S. H. (2009) Int J Clin Exp Pathol 2, 574-82
Hegazy, A. N. & Klein, C. (2008) Leukemia 22, 2070-9
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
Riera-Sans, L., & Behrens, A. (2007) J Immunol 178, 5690-700
Gerna, G., Percivalle, E., Lilleri, D., Lozza, L., Fornara, C., Hahn, G., Baldanti, F. & Revello, M. G. (2005) J Gen Virol 86, 275-84.
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
Strobl, H. & Knapp, W. (2004) J Biol Regul Homeost Agents 18, 335-9.
Haranaga, S., Yamaguchi, H., Friedman, H., Izumi, S., & Yamamoto, Y. (2001) Infect Immun 69, 7753-9
Kappelmayer, J., Gratama, J. W., Karaszi, E., Menendez, P., Ciudad, J., Rivas, R. & Orfao, A. (2000) J Immunol Methods 242, 53-65.
Millard, I., Degrave, E., Philippe, M. & Gala, J. L. (1998) Clin Chem 44, 2320-30.
Strobl, H., Scheinecker, C., Riedl, E., Csmarits, B., Bello-Fernandez, C., Pickl, W. F., Majdic, O. & Knapp, W. (1998) J Immunol 161, 740-8.
Konikova, E., Glasova, M., Kusenda, J. & Babusikova, O. (1998) Neoplasma 45, 282-91.
Millard, I., Degrave, E., Philippe, M. & Gala, J. L. (1998) Clin Chem 44, 2320-30.
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.
Sedlmayr, P., Grosshaupt, B. & Muntean, W. (1996) Cytometry 23, 284-9.
Groeneveld, K., te Marvelde, J. G., van den Beemd, M. W., Hooijkaas, H. & van Dongen, J. J. (1996) Leukemia 10, 1383-9.
Knapp, W., Strobl, H., Scheinecker, C., Bello-Fernandez, C. & Majdic, O. (1995) Ann Hematol 70, 281-96.
Scheinecker, C., Strobl, H., Fritsch, G., Csmarits, B., Krieger, O., Majdic, O. & Knapp, W. (1995) Blood 86, 4115-23.
Strobl, H., Scheinecker, C., Csmarits, B., Majdic, O. & Knapp, W. (1995) Br J Haematol 90, 774-82.
Knapp, W., Strobl, H. & Majdic, O. (1994) Cytometry 18, 187-98.
Knapp, W., Majdic, O. & Strobl, H. (1993) Recent Results Cancer Res 131, 31-40.
Strobl, H., Takimoto, M., Majdic, O., Fritsch, G., Scheinecker, C., Hocker, P. & Knapp, W. (1993) Blood 82, 2069-78.
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Product Notes
The FIX&PERM Solution A (Fix) (CE) (Catalog #AAA570407) is a Reagent and is intended for research purposes only. The product is available for immediate purchase. AAA Biotech's FIX&PERM Solution A (Fix) (CE) can be used in a range of immunoassay formats including, but not limited to, Flow Cytometry (FC/FACS). 1. For each sample to be analyzed add 50 ul of whole blood, bone marrow or mononuclear cell suspension in a 5ml tube 2. Add 100 ul of Reagent A (Fixation Medium, stored and used at room temperature) 3. Incubate for 15 minutes at room temperature 4. Add 5ml phosphate buffered saline and centrifuge cells for 5 minutes at 300g 5. Remove supernatant and add to cell pellet 100 ul Reagent B (Permeabilization Medium) and 20 ul of the appropriate monoclonal antibody conjugate 6. Vortex at low speed for 1-2 seconds 7. Incubate for 15 minutes at room temperature 8. Wash cells with phosphate buffered saline as described above 9. 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 degree 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. Researchers should empirically determine the suitability of the FIX&PERM Solution A (Fix) (CE) for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "FIX&PERM Solution A (Fix) (CE), Reagent" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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