COMBI IC Reagent: Mouse anti Myeloperoxidase-C2 (FITC) and Mouse anti Lactoferrin (PE)

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NameCOMBI IC Reagent: Mouse anti Myeloperoxidase-C2 (FITC) and Mouse anti Lactoferrin (PE)
Catalog numberGIC-212
Size50 Tests
Price€ 343.00


More details

CategoryPrimary Antibodies
Long descriptionMyeloperoxidase (MPO) is a glycoprotein present in the azurophil (primary) granules of myeloid cells, which appears in the myeloblast stage of myeloid cell differentiation. MPO is he most common functional protein of myeloid cells and is involved in the inflammatory response. It helps to kill microbes by breaking down peroxide in the presence of halide ions, contributing to the bactericidal function of granulocytes. The primary translation product of MPO undergoes glycosylation with production of the 89 kDa heme-free apopro-MPO form followed by incorporation of heme and conversion into the enzymatically active pro-MPO form. Subsequently, pro-MPO becomes targeted to azurophil granules where final processing occurs to produce mature dimeric MPO consisting of the 59-64 kDa MPO α-chain and the 14 kDa MPO β-chain. Lactoferrin (LF) is an iron-binding protein with bactericidal and bacteriostatic activity which is stored within the secondary granules of granulocytes. LF expression is restricted to the post-mitotic maturation compartment of the granulocytic lineage, starting from the myelocyte stage. Normal and malignant myeloblasts are LF negative. The combined staining for MPO and LF allows the distinction between mature and immature myelomonocytic cells. The MPO-C2/LF COMBI-IC reagent permits the identification and enumeration of immature and more mature myelomonocytic cell populations in normal and malignant human blood and bone marrow using flow cytometry. 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.
Antibody come fromn/a
Other descriptionPBS pH 7.2, 1% BSA, 0.05% NaN3
Clone8E6 and 4C5
Antigen-antibody binding interactionCOMBI IC Reagent: Mouse anti Myeloperoxidase-C2 (FITC) and Mouse anti Lactoferrin (PE) Antibody
Antibody is raised inMouse
Antibody's reacts withHuman
Antibody's reacts with these speciesThis antibody doesn't cross react with other species
Antibody's specificityAntibody MPO-C2 (clone 8E6) reacts with human myeloperoxidase (MPO) expressed by normal and malignant myelomonocytic cells. The LF mAb (clone 4C5) recognizes lactoferrin stored within secondary granules of postmitotic granulocyte-commited cells. In this COMBI-IC Reagent antibody 8E6 is conjugated to FITC, antibody 4C5 is conjugated to Phycoeythrin (PE). The sensitivity of MPO-C2/LF 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 fluorescence intensity). 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, 50 µl of leukocytes containing 10^7 cells/ml are stained with 20 µl 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 3-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.
Research interestCD Marker
ApplicationFlow cytometry
Antibody's suited forPermeabilization and Staining Procedure - In combination with our Permeabilization Kit FIX&PERM (Cat. No. GAS-002) intracellular MPO-C2 and LF can be easily stained in cell suspensions. - For each sample to be analyzed add 50 µl of whole blood, bone marrow or mononuclear cell suspension in a 5 ml tube - Add 100 µl of Reagent A (Fixation Medium, stored and used at room temperature) - Incubate for 15 minutes at room temperature - Add 5 ml phosphate buffered saline and centrifuge cells for 5 minutes at 300 g - Remove supernatant and add to cell pellet 100 µl Reagent B (Permeabilization Medium) and 20 µl of the MPO-C2/LF COMBI-IC 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
StorageNordic-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. 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.
Relevant referencesAndersson, E., Hellman, L., Gullberg, U. & Olsson, I. (1998) J Biol Chem 273, 4747-53. Braylan, R. C., Orfao, A., Borowitz, M. J. & Davis, B. H. (2001) Cytometry 46, 23-7 Catovsky, D., Matutes, E., Buccheri, V., Shetty, V., Hanslip, J., Yoshida, N. & Morilla, R. (1991) Ann Hematol 62, 16-21. Cowland, J. B. & Borregaard, N. (1999) J Leukoc Biol 66, 989-95 Groeneveld, K., te Marvelde, J. G., van den Beemd, M. W., Hooijkaas, H. & van Dongen, J. J. (1996) Leukemia 10, 1383-9 Gullberg, U., Andersson, E., Garwicz, D., Lindmark, A. & Olsson, I. (1997) Eur J Haematol 58, 137-53. He, J. & Furmanski, P. (1995) Nature 373, 721-4. Imamura, N. (1998) Am J Hematol 58, 241-3. Knapp, W., Majdic, O. & Strobl, H. (1993) Recent Results Cancer Res 131, 31-40. Koeffler, H. P., Ranyard, J. & Pertcheck, M. (1985) Blood 65, 484-91. 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. Murao, S., Stevens, F. J., Ito, A. & Huberman, E. (1988) Proc Natl Acad Sci U S A 85, 1232-6. Nakase, K., Sartor, M. & Bradstock (1998) Cytometry 34, 198-202. Nauseef, W. M. (1990) Hematol Pathol 4, 165-78. Nauseef, W. M., Olsson, I. & Arnljots, K. (1988) Eur J Haematol 40, 97-110. Oehler, L., Majdic, O., Pickl, W. F., Stockl, J., Riedl, E., Drach, J., Rappersberger, K., Geissler, K. & Knapp, W. (1998) J Exp Med 187, 1019-28. Paietta, E. (2003) Best Pract Res Clin Haematol 16, 671-83. Rado, T. A., Bollekens, J., St Laurent, G., Parker, L. & Benz, E. J., Jr. (1984) Blood 64, 1103-9. Rado, T. A., Wei, X. P. & Benz, E. J., Jr. (1987) Blood 70, 989-93. Srivastava, C. H., Rado, T. A., Bauerle, D. & Broxmeyer, H. E. (1991) J Immunol 146, 1014-9. Strobl, H. & Knapp, W. (2004) J Biol Regul Homeost Agents 18, 335-9.
Protein numbersee ncbi
WarningsFor 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.
DescriptionThis antibody needs to be stored at + 4°C in a fridge short term in a concentrated dilution. Freeze thaw will destroy a percentage in every cycle and should be avoided.
PropertiesThis nordc Fluorescein isothiocyanate (FITC) antibody is currently after some BD antibodies the most commonly used fluorescent dye for FACS. When excited at 488 nanometers, FITC has a green emission that's usually collected at 530 nanometers, the FL1 detector of a FACSCalibur or FACScan. FITC has a high quantum yield (efficiency of energy transfer from absorption to emission fluorescence) and approximately half of the absorbed photons are emitted as fluorescent light. For fluorescent microscopy applications, the 1 FITC is seldom used as it photo bleaches rather quickly though in flow cytometry applications, its photo bleaching effects are not observed due to a very brief interaction at the laser intercept. nordc FITC is highly sensitive to pH extremes.
ConjugationAnti-FITC Antibody
TestMouse or mice from the Mus musculus species are used for production of mouse monoclonal antibodies or mabs and as research model for humans in your lab. Mouse are mature after 40 days for females and 55 days for males. The female mice are pregnant only 20 days and can give birth to 10 litters of 6-8 mice a year. Transgenic, knock-out, congenic and inbread strains are known for C57BL/6, A/J, BALB/c, SCID while the CD-1 is outbred as strain.
Latin nameMus musculus