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Category
Three-Color Reagents (FITC/RPE/Cy5RPE)
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Long description
CD3-FITC/CD4-PE/CD45-PECy5 is a three-color direct immunofluorescence reagent for use in flow cytometry designed for the identification and enumeration of helper/inducer T lymphocytes (CD3+CD4+)._x000B__x000B_SUMMARY AND EXPLANATION_x000B_Flow Cytometry (FC) is a powerful tool for the analytical and quantitative characterization of cells which provides rapid, quantitative and multiparametric analysis of heterogeneous cell populations on a cell-by-cell basis. Flow cytometry is performed on cells in liquid suspension that have been incubated with fluorescently-labeled antibodies directed against specific cellular proteins. The relative fluorescence intensity of the positive cells indicates the amount of antibody bound to specific binding sites on the cells, and therefore provides a relative measure of antigen expression._x000B__x000B_Human lymphocytes may be classified in three main populations according to their biological function and their cell surface antigen expression: T lymphocytes, B lymphocytes and natural killer cells (NK). T lymphocytes (CD3+), the precursors of which originate in the bone marrow and then migrate and mature in the thymus, can be subdivided as well in functionally different populations. The most clearly defined of these are helper/inducer T cells (CD3+CD4+) and suppressor/cytotoxic T cells (CD3+CD8+). T cells produce no antibodies and are the mediators of cell immunity._x000B_The CD3-FITC/CD4-PE/CD45-PECy5 reagent reco_x000B_gnizes the antigen CD3 present in T lymphocytes, the antigen CD4 present in helper/inducer T cells (CD3+CD4+) and the antigen CD45, the leucocyte common antigen (LCA) complex, which is expressed exclusively on cells of the haematopoietic system (lymphocytes, monocytes, and granulocytes) and their progenitors. This reagent can be used in the characterization studies for immunophenotyping of lymphocytes, which are widely applied for monitoring of the immunologic status of post-transplant patients and in the characterization and follow-up of immunodeficiencies, autoimmune diseases, leukemia etc (1-2). Individuals with HIV typically exhibit a steady decrease of helper/inducer T lymphocyte counts as the infection progresses. The CD4 antigen is also detected on some monocytes (CD3-CD4+)._x000B__x000B_PRINCIPLES OF THE PROCEDURE_x000B_Flow cytometry is an innovative technology by means of which different cell characteristics are simultaneously analyzed on a single cell basis. This is achieved by means of hydrodynamic focusing of cells that pass aligned one by one in front of a set of light detectors; at the same time they are illuminated by a laser beam. The interaction of the cells with the laser beam generates signals of two different kinds: those generated by dispersed light (FSC/SSC), which mainly reflects the size of the cell and its internal complexity, and those related to the emission of light by the fluorochromes present in the cell. These signals become electric impulses which are amplified and registered as digital signals to be processed by a computer._x000B__x000B_When the reagent is added to the sample, the fluorochrome-labelled monoclonal antibodies (MAb) present in the reagent (CD3-FITC, CD4-PE and CD45-PECy5) bind specifically to the antigens they are directed against, allowing the detection by FC of the cell populations carried by the antigens. The erythrocyte population, which could hinder the detection of the target population, is eliminated by the use of a red blood cell lysing solution previous to acquire the sample on the cytometer. The use of QuicklysisTM (CYT-QL-1) erythrocyte lysing solution is recommended, since it requires no further washing step and contains no fixative, therefore minimizing the handling of the sample and avoiding the cell loss associated to the centrifuge process.(3,4)._x000B__x000B_The counts of helper/inducer T cells (CD3+CD4+) are expressed as a percentage of the total amount of lymphocytes or leucocytes present in the sample which can itself be determined by FC based on its typical pattern of SSC / CD45+. Because each flow cytometer has different operating characteristics each laboratory must determine its optimal operating procedure.
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Antibody come from
CD3=Derived from the hybridization of mouse NS-1 myeloma celss with spleen cells from BALB/c mice immunized with human thymocytes. CD4=Derived from the hybridization of mouse NS-1 myeloma cells with spleen cells from BALB/c mice immunized with human perherial blood T lymphocytes.
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Other description
Provided as solution in phosphate buffered saline with 0.1% sodium azide. Affinity chromatography
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Clone
not specified
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Antigen antibody binding interaction
Mouse anti CD3 antibody conjugated to FITC / CD4 antibody conjugated to PE / CD45 antibody conjugated to Cy5-PE Antibody
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Antibody is raised in
Mouse
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Antibody s reacts with
Human
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Antibody s reacts with these species
This antibody doesn't cross react with other species
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Antibody s specificity
No Data Available
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Research interest
CD Marker
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Application
Flow Cytometry
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Antibody s suited for
WARNINGS AND RECOMMENDATIONS_x000B_1.For in vitro diagnostic use. _x000B_2.This product is supplied ready to use. If it is altered by dilution or addition of other components, it will be invalidated for in vitro_x000B_diagnostic use. _x000B_3.The reagent is stable until the expiration date shown on the label if it is properly stored. Do not use after the expiration date shown_x000B_on the label. If the reagents are stored in conditions different from those recommended, such conditions must be validated by the_x000B_user. _x000B_4.Alteration in the appearance of the reagent, such as the precipitation or discoloration indicates instability or deterioration. In such_x000B_cases, the reagent should not be used. _x000B_5.It contains 0.1% sodium azide (CAS-Nr. 26628-22-8) as a preservative, but even so care should be taken to avoid microbial_x000B_contamination of reagent or incorrect results may occur. _x000B_¥Sodium azide (NaN3) is harmful if swallowed (R22), if swallowed, seek medical advice immediately and show this_x000B_container or label (S46)._x000B_¥Wear suitable protecting clothing (S36). _x000B_¥Contact with acids liberates very toxic gas (R32). _x000B_¥Azide compounds should be flushed with large volumes of water during disposal to avoid deposits in metal drains where explosive conditions may develop._x000B_6.All patient specimens and materials with which they come into contact are considered biohazards and should be handled as if capable of transmitting infection (5), and disposed according to the legal precautions established for this type of product. Also recommended is handling of the product with appropriate protective gloves and clothing, and its use by personnel sufficiently qualified for the procedures described. Avoid contact of samples with skin and mucous membranes. After contact with skin, wash immediately with plenty of water._x000B_7.Use of the reagent with incubation times or temperatures different from those recommended may cause erroneous results. Any such changes must be validated by the user._x000B__x000B_WARRANTY_x000B_This product is warranted only to conform to the quantity and contents stated on the label. There are no warranties that extend beyond the description on the label of the product. CytognosÕs sole liability is limited to either replacement of the product or refund of the purchase price.
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Storage
4ºC
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Relevant references
1.Orfao A, Gonz‡lez de Buitrago JM La citometr’a de flujo en el laboratorio cl’nico. Sociedad espa–ola de bioqu’mica cl’nica y patolog’a Molecular 1995._x000B_2. Stetler-Stevenson M. Flow cytometry analysis of lymphomas and lymphoproliferative disorders. Semin Hematol 2001 Apr;38(2):111-23._x000B_3.MenŽndez P, et al. Comparison between a lyse-and-then-wash method and a lyse-non-wash technique for the enumeration of CD34+ hematopoietic progenitor cells. Cytometry (Comm. Clin. Cytometry) 34: 264-271 (1998)_x000B_4.GratamaJW, MenŽndez P, Kraan J, Orfao A. Loss of CD34+ hematopoietic progenitor cells due to washing can be reduced by the use of fixative-free erytrocyte lysing reagents. J Immunol. Methods 239: 13-23 (2000)_x000B_5.Protection of Laboratory Workers from occupationally acquired infections. Second edition; approved guideline (2001). Villanova PA: National Committee for Clinical Laboratory Standards; Document M29-A2._x000B_6.Procedures for the collection of diagnostic blood specimens by venipuncture- approved standard; Fifth edition (2003). Wayne PA: National Committee for Clinical Laboratory Standards; Document H3-A5._x000B_7.Clinical applications of flow cytometry: Quality assurance and immunophenotyping of lymphocytes; approved guideline (1998). Wayne PA: National Committee for Clinical Laboratory Standards; Document H42-A._x000B_8.Lima M et al. TCR +/CD4+ Large Granular Lymphocytosis. A new Clonal T-Cell Lymphoprolipherative Disorder. American Journal of Pathology, 163 (2):763-771 (2003)_x000B_9.Gorczyza W. et al. An approach to diagnosis of T-cell lymphoprolipherative disorders by flow cytometry. Cytometry (Clinical cytometry) 50:177-190 (2002)_x000B_10. Braylan RC, Orfao A, Borowitz MJ, Davis BH. Optimal number of reagents required to evaluate hematolymphoid neoplasias: results of an international consensus meeting. Cytometry 46: 23-7 (2001)_x000B_11.Jennings CD, Foon KA. Recent advances in flow cytometry: application to the diagnosis of hematologic malignancy. Blood 90(8): 2863-2892 (1997)_x000B_12.Reichert et al. Lymphocyte subset reference ranges in adult Caucasians. Clin Immunol Immunopathol 60:190-208 (1991) _x000B_13.Prince HK et al. Influence of racial background on the distribution of T-cell subsets and Leu-11 positive lymphocytes in healthy_x000B_blood donors. Diagn Immunol. 3: 33-39 (1985) _x000B_14.Kotylo PK et al. Reference ranges for lymphocyte subsets in pediatric patients. Am J Clin Pathol 100:111-5 (1993)
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Protein number
see ncbi
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Warnings
This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving humans or animals. This datasheet is as accurate as reasonably achievable, but Nordic-MUbio accepts no liability for any inaccuracies or omissions in this information.
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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. CD4+ T helper cell marker or cluster differentiator of white blood cells in FACS or flow cytometry. Used offer coupled to magnetic beads. GENTAUR makes custom magnetic bead couplings on 50 and 250 nm beads for certain clones.
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Properties
If you buy Antibodies supplied by nordc they should be stored frozen at - 24°C for long term storage and for short term at + 5°C. Cy5 antibodies are excited by the 650-nanometer wave of an argon laser and detected at a 670-nanometer captor. 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.
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Conjugation
Cy5.5, Anti-FITC Antibody
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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.
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Latin name
Mus musculus
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French translation
anticorps
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Gene target
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Gene symbol
CD4, PTPRCAP, PTPRC
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Short name
Mouse anti CD3 antibody conjugated FITC / CD4 antibody conjugated PE / CD45 antibody conjugated -PE
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Technique
Antibody, Mouse, anti, FITC, antibody to, antibodies against human proteins, antibodies for, antibody Conjugates, Fluorescein, mouses
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Host
mouse
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Isotype
not specified
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Label
FITC and PE and Cy5-PE
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Species
Mouse, Mouses
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Alternative name
Mouse antibody to CD3 (antibody to-) coupled to fluorecein / CD4 molecule (antibody to-) coupled to peroxidase / CD45 (antibody to-) coupled to cyanine 5-peroxidase
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Alternative technique
antibodies, murine, fluorescine
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Alternative to gene target
CD4 molecule, CD4mut, CD4 and IDBG-15034 and ENSG00000010610 and 920, protein homodimerization activity, Cell surfaces, Cd4 and IDBG-188351 and ENSMUSG00000023274 and 12504, CD4 and IDBG-640247 and ENSBTAG00000003255 and 407098
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