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V500 Rat anti-Mouse CD4
V500 Rat anti-Mouse CD4

Analysis of CD4 on mouse splenocytes. Splenocytes from BALB/c mice were stained simultaneously with BD Horizon™ V500 Rat anti-Mouse CD4 (right panel) or BD Horizon™ V500 Rat IgG2a, κ Isotype Control (clone R35-95, Cat. No. 560786, left panel), and PE Hamster Anti-Mouse CD3e (clone 145-2C11, Cat. No. 553063/553064). The contour plots were derived from gated events based on light scattering characteristics of splenocytes. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.

Analysis of CD4 on mouse splenocytes. Splenocytes from BALB/c mice were stained simultaneously with BD Horizon™ V500 Rat anti-Mouse CD4 (right panel) or BD Horizon™ V500 Rat IgG2a, κ Isotype Control (clone R35-95, Cat. No. 560786, left panel), and PE Hamster Anti-Mouse CD3e (clone 145-2C11, Cat. No. 553063/553064). The contour plots were derived from gated events based on light scattering characteristics of splenocytes. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.

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 DA, also known as DA/HA IgG2a, κ
Mouse Thymocytes (BALB/c)
Flow cytometry (Routinely Tested)
0.2 mg/ml
12504
AB_1937327
Aqueous buffered solution containing protein stabilizer, glycerol and ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with BD Horizon™ V500 under optimum conditions, and unreacted BD Horizon™ V500 was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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. BD Horizon V500 has a maximum absorption of 415 nm and maximum emission of 500 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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Antibody Details
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RM4-5

The RM4-5 monoclonal antibody specifically binds to the CD4 (L3T4) differentiation antigen expressed on most thymocytes, subpopulations of mature T lymphocytes (i.e., MHC class II-restricted T cells, including most T helper cells and immunosuppressive regulatory T cells), and a subset of NK-T cells. CD4 has also been reported to be detected on pluripotent hematopoietic stem cells, bone marrow myeloid and B-lymphocyte precursors, intrathymic lymphoid precursors, and a subset of splenic dendritic cells. CD4 has 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. Purified RM4-5 mAb has been reported to block the binding of FITC-conjugated anti-mouse CD4 clones GK1.5 and H129.19, but not the RM4-4 clone.

The antibody is conjugated to BD Horizon™ V500, which has been developed for use in multicolor flow cytometry experiments and is

available exclusively from BD Biosciences. It is excited by the Violet laser with an Ex max of 415 nm and Em Max at 500 nm. BD Horizon V500 conjugates emit at a similar wavelength to Amcyan yet exhibit reduced spillover into the FITC channel. For more information on BD Horizon V500, visit bdbiosciences.com/colors.

When compensating dyes in this spectral range (such as Horizon™ V500 and AmCyan), the most accurate compensation can be obtained using single stained cellular controls.  Due to spectral differences between cells and beads  in this channel, using BD CompBeads can result in spillover errors for V500 and AmCyan reagents.   Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended.   Different V500 reagents (e.g. CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone specific compensation controls when using these reagents.

560782 Rev. 2
Format Details
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V500
BD Horizon™ V500 Dye is part of the BD Horizon™ Violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 415-nm and an emission maximum (Em Max) at 499-nm. Driven by BD innovation, V500 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 500 nm (eg, a 525/50 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
V500
Violet 405 nm
415 nm
499 nm
560782 Rev.2
Citations & References
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Development References (13)

  1. Allman D, Li J, Hardy RR. Commitment to the B lymphoid lineage occurs before DH-JH recombination. J Exp Med. 1999; 189(4):735-740. (Biology). View Reference
  2. Bendelac A. Mouse NK1+ T cells. Curr Opin Immunol. 1995; 7(3):367-374. (Biology). View Reference
  3. Bierer BE, Sleckman BP, Ratnofsky SE, Burakoff SJ. The biologic roles of CD2, CD4, and CD8 in T-cell activation. Annu Rev Immunol. 1989; 7:579-599. (Biology). View Reference
  4. 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. (Biology). View Reference
  5. Godfrey DI, Kennedy J, Mombaerts P, Tonegawa S, Zlotnik A. Onset of TCR-β gene rearrangement and role of TCR-β expression during CD3-CD4-CD8- thymocyte differentiation. J Immunol. 1994; 152(10):4783-4792. (Biology). View Reference
  6. Guo MW, Watanabe T, Mori E, Mori T. Molecular structure and function of CD4 on murine egg plasma membrane. Zygote. 1995; 3(1):65-73. (Biology). View Reference
  7. 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
  8. Martin P, del Hoyo GM, Anjuere F, et al. Concept of lymphoid versus myeloid dendritic cell lineages revisited: both CD8alpha(-) and CD8alpha(+) dendritic cells are generated from CD4(low) lymphoid-committed precursors. Blood. 2000; 96(7):2511-2519. (Biology). View Reference
  9. Nakamura T. Personal Communication. .
  10. Shevach EM. Regulatory T cells in autoimmmunity. Annu Rev Immunol. 2000; 18:423-449. (Biology). View Reference
  11. 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. (Biology). View Reference
  12. 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
  13. Wu L, Scollay R, Egerton M, Pearse M, Spangrude GJ, Shortman K. CD4 expressed on earliest T-lineage precursor cells in the adult murine thymus. Nature. 1991; 349(6304):71-74. (Biology). View Reference
View All (13) View Less
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