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PE-CF594 Mouse Anti-Human CD4
PE-CF594 Mouse Anti-Human CD4
Multiparameter flow cytometric analysis of CD4 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ PE-CF594 Mouse IgG2b, κ Isotype Control (Cat. No. 562305; Left Plot) or BD Horizon PE-CF594 Mouse Anti-Human CD4 antibody (Cat. No. 566707/566709; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The two-parameter pseudocolor density plot showing the correlated expression of CD4 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of CD4 expression on human peripheral blood leucocyte populations. Human whole blood was stained with either BD Horizon™ PE-CF594 Mouse IgG2b, κ Isotype Control (Cat. No. 562305; Left Plot) or BD Horizon PE-CF594 Mouse Anti-Human CD4 antibody (Cat. No. 566707/566709; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The two-parameter pseudocolor density plot showing the correlated expression of CD4 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of intact leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Product Details
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BD Horizon™
CD4 antigen (p55); T-cell surface antigen T4/Leu-3
Human (QC Testing)
Mouse BALB/c x A/J, also known as CAF1 IgG2b, κ
Sheep Erythrocyte Rosette-purified Human T Cells
Flow cytometry (Routinely Tested)
5 µl
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unconjugated antibody and free dye were removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation).  When fluorochrome conjugated antibodies are bound to 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 cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. 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).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. An isotype control should be used at the same concentration as the antibody of interest.
  8. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  9. CF™ is a trademark of Biotium, Inc.
  10. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  11. This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  12. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
  13. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
566707 Rev. 1
Antibody Details
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OKT4

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.

This antibody is conjugated to BD Horizon™ PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg, 610/20-nm filter).

566707 Rev. 1
Format Details
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PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, is designed to be excited by the blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
566707 Rev.1
Citations & References
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Development References (9)

  1. Courtney AH, Lo WL, Weiss A. TCR Signaling: Mechanisms of Initiation and Propagation. Trends Biochem Sci. 2018; 43(2):108-123. (Biology). View Reference
  2. 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
  3. 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.
  4. Kung P, Goldstein G, Reinherz EL, Schlossman SF. Monoclonal antibodies defining distinctive human T cell surface antigens. Science. 1979; 206(4416):347-349. (Immunogen: Cytotoxicity, Flow cytometry, Radioimmunoassay). View Reference
  5. 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.
  6. 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.
  7. 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
  8. 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):2894-2896. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  9. 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):4061-4065. (Immunogen: Cell separation, Flow cytometry, Fluorescence activated cell sorting). View Reference
View All (9) View Less
566707 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims.  Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.