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Multiparameter flow cytometric analysis of CD4 expression on Human peripheral blood leukocyte populations. Human whole blood was stained with either FITC Mouse IgG2b, κ Isotype Control (Cat. No. 565379; Left Plot) or FITC Mouse Anti-Human CD4 antibody (Cat. No. 566802; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The two-parameter pseudocolor dot plot showing the correlated expression of CD4 (or Ig Isotype control staining) versus side light-scatter (SSC) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocyte populations. Flow cytometry and data analysis were performed using a BD FACSCelesta™ Flow Cytometer System and FlowJo™ Software.
BD Pharmingen™ FITC Mouse Anti-Human CD4
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Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
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Companion Products
The OKT4 monoclonal antibody specifically recognizes CD4 which is also known as T4 and Leu-3. CD4 is expressed on most thymocytes, T-helper/inducer cells, regulatory T cells (Tregs), and NKT cells. It is also variably expressed on monocytes, macrophages, and some dendritic cells. CD4 is a ~55 kDa type I transmembrane glycoprotein that is encoded by CD4 which belongs to the immunoglobulin superfamily (IgSF). CD4 binds to a nonpolymorphic region of MHC class II and serves as a coreceptor, along with the T cell receptor for antigen (TCR), for the MHC Class II-restricted recognition of antigens on antigen-presenting cells. CD4 also functions as a receptor for human immunodeficiency viruses (HIV). CD4 has four immunoglobulin-like extracellular domains in its extracellular region and a CXCP binding motif for p56Lck in its cytoplasmic domain which regulates the antigen-bound TCR-induced signaling cascade.
Development References (9)
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Courtney AH, Lo WL, Weiss A. TCR Signaling: Mechanisms of Initiation and Propagation.. Trends Biochem Sci. 2018; 43(2):108-123. (Biology). View Reference
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Hoffman RA, Kung PC, Hansen WP, Goldstein G.. Simple and rapid measurement of human T lymphocytes and their subclasses in peripheral blood.. Proc Natl Acad Sci U S A. 1980; 77(8):4914-4917. (Clone-specific: Flow cytometry). View Reference
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Horibe K, Knowles RW, Naito K, Morishima Y, Dupont B. Analysis of T lymphocyte antibody specificities: Comparison of serology with immunoprecipitation patterns. In: Bernard A. A. Bernard .. et al., ed. Leucocyte typing : human leucocyte differentiation antigens detected by monoclonal antibodies. Berlin New York: Springer-Verlag; 1984:212-224.
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Kung P, Goldstein G, Reinherz EL, Schlossman SF. Monoclonal antibodies defining distinctive human T cell surface antigens.. Science. 1979; 206(4416):347-9. (Immunogen: Cytotoxicity, Flow cytometry, Radioimmunoassay). View Reference
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Miedema F, Terpstra FG, Melief CJM. T Cell-dependent immunoglobulin synthesis in the human system. Studies with T cell-specific monoclonal antibodies. In: Reinherz EL. Ellis L. Reinherz .. et al., ed. Leukocyte typing II. New York: Springer-Verlag; 1986:213-222.
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Moebius U. Cluster report: CD4. In: Knapp W. W. Knapp .. et al., ed. Leucocyte typing IV : white cell differentiation antigens. Oxford New York: Oxford University Press; 1989:314-330.
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Reinherz EL, Kung PC, Goldstein G, Levey RH, Schlossman SF. Discrete stages of human intrathymic differentiation: analysis of normal thymocytes and leukemic lymphoblasts of T-cell lineage. Proc Natl Acad Sci U S A. 1980; 77(3):1588-1592. (Clone-specific: Cell separation, Cytotoxicity, Flow cytometry). View Reference
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Reinherz EL, Kung PC, Goldstein G, Schlossman SF. Further characterization of the human inducer T cell subset defined by monoclonal antibody.. J Immunol. 1979; 123(6):2894-6. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
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Reinherz EL, Kung PC, Goldstein G, Schlossman SF. Separation of functional subsets of human T cells by a monoclonal antibody.. Proc Natl Acad Sci U S A. 1979; 76(8):4061-5. (Immunogen: Cell separation, Flow cytometry, Fluorescence activated cell sorting). View Reference
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