COMBI IC Reagent: Mouse anti CD45 (FITC) and Mouse anti CD14 (PE)

  • Catalog number
    GCT-201
  • Price
    Please ask
  • Size
    50 Tests
  • Category
    Primary Antibodies
  • Long description
    The CD45 molecule is typically expressed at high levels on all hematopoietic cells. CD45 is a major component of the glycocalix of these cells and can be expressed in different isoforms. Antibody VIT200 recognizes a pan CD45 epitope, which is expressed on all hematopoietic cells. CD14 is a GPI-anchored molecule expressed by virtually all human monocytes and macrophages and – to a lesser degree - granulocytes. CD14 together with Toll-like receptor 4 and MD-2 forms the LPS-receptor complex that recognizes and signals the presence of LPS. While CD14 has no signaling structure its main role seems to be the binding of LPS. The VIT200/MEM18 COMBI-REAGENT-Gate Control permits the identification and enumeration of human leukocytes 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 from
    n/a
  • Other description
    PBS pH 7.2, 1% BSA, 0.05% NaN3
  • Clone
    VIT200 and MEM18
  • Antigen antibody binding interaction
    COMBI IC Reagent: Mouse anti CD45 (FITC) and Mouse anti CD14 (PE) Antibody
  • Antibody is raised in
    Mouse
  • Antibody s reacts with
    Human
  • Antibody s reacts with these species
    This antibody doesn't cross react with other species
  • Antibody s specificity
    The CD45 mAb (clone VIT200) recognizes a pan CD45 epitope. The CD14 mAb (clone MEM18) recognizes surface CD14 on human monocytes and macrophages as well as neutrophils. The sensitivity of CD45/CD14 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 interest
    CD Marker
  • Application
    Direct Immunofluorescence, Flow Cytometry
  • Antibody s suited for
    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 conjugate - 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 - Centrifuge 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 conjugate 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 - Centrifuge tubes for 5 minutes at 300 g - Remove supernatant, resuspend cells in 2-5 ml of phosphate buffered saline (PBS) and centrifuge 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
  • Storage
    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. 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 references
    Battifora, H. & Trowbridge, I. S. (1983) Cancer 51, 816-21. Beutler, B. (2002) Curr Top Microbiol Immunol 270, 109-20. Brocklebank, A. M. & Sparrow, R. L. (2001) Cytometry 46, 254-61. Cobbold, S., Hale, G. & Waldmann, H. (1987) Leucocyte Typing III. p788-803 (Oxford University Press, Oxford-New York-Tokyo). Dalchau, R., Kirkley, J. & Fabre, J. W. (1980) Eur J Immunol 10, 737-44. Goyert, S. M. (1989) Leucocyte Typing IV. p789-793 (Oxford University Press, Oxford-New York-Tokyo). Goyert, S. M., Ferrero, E., Rettig, W. J., Yenamandra, A. K., Obata, F. & Le Beau, M. M. (1988) Science 239, 497-500. Goyert, S. M., Ferrero, E. M., Seremetis, S. V., Winchester, R. J., Silver, J. & Mattison, A. C. (1986) J Immunol 137, 3909-14. Jing, S., Ralph, S., Head, M. T. A., Chain, A. & Trowbridge, I. (1987) Structural studies of the transferrin receptor and T 200 glycoprotein (CD45). Leucocyte Typing III. p899-905 (Oxford University Press, Oxford-New York-Tokyo). Knapp, W. (1982) Blut 45, 301-8. Knapp, W. (1989) Leucocyte Typing IV. p747-780 (Oxford University Press, Oxford-New York-Tokyo). Means, T. K., Lien, E., Yoshimura, A., Wang, S., Golenbock, D. T. & Fenton, M. J. (1999) J Immunol 163, 6748-55. Nicholson, J. K., Hubbard, M. & Jones, B. M. (1996) Cytometry 26, 16-21. Sugita, K., Majdic, O., Stockinger, H., Holter, W., Burger, R. & Knapp, W. (1987) Transplantation 43, 570-4. Sun, T., Sangaline, R., Ryder, J., Gibbens, K., Rollo, C., Stewart, S. & Rajagopalan, C. (1997) Am J Clin Pathol 108, 152-7. Tapping, R. I., Akashi, S., Miyake, K., Godowski, P. J. & Tobias, P. S. (2000) J Immunol 165, 5780-7. Thomas, M. L. (1989) Annu Rev Immunol 7, 339-69. Ugolini, V., Nunez, G., Smith, R. G., Stastny, P. & Capra, J. D. (1980) Proc Natl Acad Sci U S A 77, 6764-8. Yoshimura, A., Lien, E., Ingalls, R. R., Tuomanen, E., Dziarski, R. & Golenbock, D. (1999) J Immunol 163, 1-5.
  • Protein number
    see ncbi
  • Warnings
    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.
  • Description
    This 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.
  • Properties
    This 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.
  • Conjugation
    Anti-FITC Antibody
  • Test
    Mouse 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 name
    Mus musculus
  • Gene target
    COMBI   Reagent   CD45   CD14  
  • Gene symbol
    CD14, PTPRCAP, PTPRC
  • Short name
    COMBI IC Reagent: Mouse anti CD45 (FITC) Mouse anti CD14 (PE)
  • Technique
    Mouse, anti, FITC, antibody to, Fluorescein, mouses
  • Host
    mouse
  • Isotype
    IgG2a and IgG1
  • Label
    FITC and PE
  • Species
    Mouse, Mouses
  • Alternative name
    COMBI IC Reagent: Mouse antibody to CD45 (fluorecein) and Mouse antibody to CD14 molecule (peroxidase)
  • Alternative technique
    murine, antibodies, fluorescine
  • Alternative to gene target
    CD14 molecule, CD14 and IDBG-49157 and ENSG00000170458 and 929, lipoteichoic acid binding, Extracellular, Cd14 and IDBG-138525 and ENSMUSG00000051439 and 12475, CD14 and IDBG-646064 and ENSBTAG00000015032 and 281048
Gene info
Gene info
Gene info
MeSH Data
  • Name
  • Concept
    Scope note: Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes.
  • Tree numbers
    • E05.196.401.143
    • E05.301.300.096
    • E05.478.566.320.200
    • E05.601.262
    • E05.601.470.320.200
  • Qualifiers
    ethics, trends, veterinary, history, classification, economics, instrumentation, methods, standards, statistics & numerical data
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