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R718 Mouse Anti-Human CD4
R718 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™ R718 Mouse IgG2b, κ Isotype Control (Cat. No. 566952; Left Plot) or BD Horizon™ R718 Mouse Anti-Human CD4 antibody (Cat. No. 567349/567350; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). A bivariate 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™ X-20 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™ R718 Mouse IgG2b, κ Isotype Control (Cat. No. 566952; Left Plot) or BD Horizon™ R718 Mouse Anti-Human CD4 antibody (Cat. No. 567349/567350; Right Plot). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). A bivariate 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™ X-20 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 ≤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 unreacted dye was removed.

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 cells and BD CompBead to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. 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).
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. This product is provided under an Agreement between BIOTIUM and BD Biosciences. 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.
  7. Alexa Fluor™ is a trademark of Life Technologies Corporation.
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.

The antibody was conjugated to BD Horizon™ Red 718, which has been developed exclusively by for BD Biosciences as a better alternative to Alexa Fluor™ 700. BD Horizon Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor™ 700.

Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
Citations & References
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View product citations for antibody "567350" on CiteAb

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

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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.