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RY586 Rat Anti-Mouse CD4
RY586 Rat Anti-Mouse CD4
Multicolor flow cytometric analysis of CD4 expression on mouse splenocytes. Splenic leucocytes from a BALB/c mouse were stained with BD Horizon™ BV421 Hamster Anti-Mouse CD3e antibody (Cat. No. 562600) and with either BD Horizon™ RY586 Rat IgG2b, κ Isotype Control (Cat. No. 568160; Left Plot) or BD Horizon™ RY586 Rat Anti-Mouse CD4 antibody (Cat. No. 568161/568162; Right Plot) at 0.5 µg/test. The bivariate pseudocolor density plot showing the correlated expression of CD4 (or Ig Isotype control staining) versus CD3 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 FACSymphony™ A5 SE Flow Cytometer and FlowJo™ software.​ Data shown on this Technical Data Sheet are not lot specific.
Multicolor flow cytometric analysis of CD4 expression on mouse splenocytes. Splenic leucocytes from a BALB/c mouse were stained with BD Horizon™ BV421 Hamster Anti-Mouse CD3e antibody (Cat. No. 562600) and with either BD Horizon™ RY586 Rat IgG2b, κ Isotype Control (Cat. No. 568160; Left Plot) or BD Horizon™ RY586 Rat Anti-Mouse CD4 antibody (Cat. No. 568161/568162; Right Plot) at 0.5 µg/test. The bivariate pseudocolor density plot showing the correlated expression of CD4 (or Ig Isotype control staining) versus CD3 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 FACSymphony™ A5 SE Flow Cytometer and FlowJo™ software.​ Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Horizon™
Cd4; CD4 antigen; L3T4; Ly-4; T-cell surface antigen T4/Leu-3
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2b, κ
Mouse CTL clone V4
Flow cytometry (Routinely Tested)
0.2 mg/ml
12504
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® CompBeads 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. An isotype control should be used at the same concentration as the antibody of interest.
  4. CF™ is a trademark of Biotium, Inc.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
568162 Rev. 2
Antibody Details
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GK1.5

The GK1.5 monoclonal antibody specifically binds to the mouse CD4 (L3T4) differentiation antigen. CD4 is expressed on most thymocytes, a subpopulation of mature T lymphocytes (i.e., MHC class II-restricted T cells, including most T helper cells), and a subset of NK-T cells. In addition, CD4 has also been reported to be detectable on pluripotent hematopoietic stem cells, bone marrow myeloid and B-lymphocyte precursors, intrathymic lymphoid precursors, and a subset of splenic dendritic cells. CD4 has also been reported to be expressed on the plasma membrane of mouse egg cells and is involved in adhesion of the egg to MHC class II-bearing sperm.  CD4 is an antigen coreceptor on the T-cell surface which interacts with MHC class II molecules on antigen-presenting cells. It participates in T-cell activation through its association with the T-cell receptor complex and protein tyrosine kinase lck. The GK1.5 antibody reportedly blocks binding of the RM4-5 and H129.19, but not RM4-4 mouse CD4-specific antibodies.

The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.

568162 Rev. 2
Format Details
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RY586
The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
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RY586
Yellow-Green 561 nm
564 nm
586 nm
568162 Rev.2
Citations & References
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View product citations for antibody "568162" on CiteAb

Development References (10)

  1. Bosselut R, Zhang W, Ashe JM, Kopacz JL, Samelson LE, Singer A. Association of the adaptor molecule LAT with CD4 and CD8 coreceptors identifies a new coreceptor function in T cell receptor signal transduction. J Exp Med. 1999; 190(10):1517-1526. (Clone-specific: (Co)-stimulation, Immunoprecipitation). View Reference
  2. Dialynas DP, Quan ZS, Wall KA, et al. Characterization of the murine T cell surface molecule, designated L3T4, identified by monoclonal antibody GK1.5: similarity of L3T4 to the human Leu-3/T4 molecule. J Immunol. 1983; 131(5):2445-2451. (Immunogen: Blocking, Depletion, Flow cytometry). View Reference
  3. Dialynas DP, Wilde DB, Marrack P, et al. Characterization of the murine antigenic determinant, designated L3T4a, recognized by monoclonal antibody GK1.5: expression of L3T4a by functional T cell clones appears to correlate primarily with class II MHC antigen-reactivity. Immunol Rev. 1983; 74:29-56. (Clone-specific: Blocking, Depletion, Flow cytometry). View Reference
  4. Frederickson GG, Basch RS. L3T4 antigen expression by hemopoietic precursor cells. J Exp Med. 1989; 169(4):1473-1478. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  5. Ghobrial RR, Boublik M, Winn HJ, Auchincloss H Jr. In vivo use of monoclonal antibodies against murine T cell antigens. Clin Immunol Immunopathol. 1989; 52(3):486-506. (Biology). View Reference
  6. Janeway CA Jr. The T cell receptor as a multicomponent signalling machine: CD4/CD8 coreceptors and CD45 in T cell activation. Annu Rev Immunol. 1992; 10:645-674. (Biology). View Reference
  7. Kruisbeek AM. In vivo depletion of CD4- and CD8-specific T cells. Curr Protoc Immunol. 1991; 4.1.1-4.1.5. (Clone-specific: Cytotoxicity, Depletion). View Reference
  8. Wineman JP, Gilmore GL, Gritzmacher C, Torbett BE, Muller-Sieburg CE. CD4 is expressed on murine pluripotent hematopoietic stem cells. Blood. 1992; 180(7):1717-1724. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting, Immunofluorescence, Immunoprecipitation). View Reference
  9. Wu L, Antica M, Johnson GR, Scollay R, Shortman K. Developmental potential of the earliest precursor cells from the adult mouse thymus. J Exp Med. 1991; 174(6):1617-1627. (Biology). View Reference
  10. Zheng B, Han S, Kelsoe G. T helper cells in murine germinal centers are antigen-specific emigrants that downregulate Thy-1. J Exp Med. 1996; 184(3):1083-1091. (Clone-specific: Immunohistochemistry). View Reference
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568162 Rev. 2

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