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BV421 Mouse Anti-Human CD4
BV421 Mouse Anti-Human CD4
Multiparameter flow cytometric analysis of CD4 expression on human peripheral blood leucocyte populations. Whole blood was stained with either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748; Left Plot) or BD Horizon BV421 antibody (Cat. No. 566703/566706; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A 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 viable 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. Whole blood was stained with either BD Horizon™ BV421 Mouse IgG2b, κ Isotype Control (Cat. No. 562748; Left Plot) or BD Horizon BV421 antibody (Cat. No. 566703/566706; Right Plot). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). A 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 viable 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
AB_2869825
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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 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.

     For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

  1. 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).
  2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  3. 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.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566703 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.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10-fold improvement in brightness compared to Pacific Blue™ conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue, and BD Horizon V450 cannot be used simultaneously.

566703 Rev. 1
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
566703 Rev.1
Citations & References
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View product citations for antibody "566703" 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
566703 Rev. 1

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