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V500 Mouse Anti-Human CD3
V500 Mouse Anti-Human CD3
Analysis of CD3 on peripheral blood lymphocytes of rhesus macaque (Macaca mulatta) monkey. Peripheral blood lymphocytes were stained with BD Horizon™ V500 Mouse Anti-Human CD3 (Cat. No. 560772/560770; solid line histogram) and compared to whole blood stained with BD Horizon™ V500 Mouse IgG1, κ Isotype Control (Cat. No. 560787; dotted line histogram). Lymphocytes were selected by light scatter profile. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
Analysis of CD3 on peripheral blood lymphocytes of rhesus macaque (Macaca mulatta) monkey. Peripheral blood lymphocytes were stained with BD Horizon™ V500 Mouse Anti-Human CD3 (Cat. No. 560772/560770; solid line histogram) and compared to whole blood stained with BD Horizon™ V500 Mouse IgG1, κ Isotype Control (Cat. No. 560787; dotted line histogram). Lymphocytes were selected by light scatter profile. Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
製品詳細
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BD Horizon™
CD3E; CD3-epsilon; T3E; TCRE
Rhesus, Cynomolgus, Baboon (QC Testing), Human (Tested in Development)
Mouse BALB/c IgG1, λ
Purified Human CD3ε Protein
Flow cytometry (Routinely Tested)
5 µl
Aqueous buffered solution containing protein stabilizer 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™ V500 under optimum conditions, and unreacted BD Horizon™ V500 was removed.

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. An isotype control should be used at the same concentration as the antibody of interest.
  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.
  6. Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
  7. BD Horizon V500 is covered by one or more of the following US patents: 8,431,416.
  8. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560772 Rev. 2
抗体の詳細
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SP34-2

Clone SP34-2 is a mouse IgG1 isotype monoclonal antibody, descendant of SP34 (mouse IgG3), with the same specificity and reactivity pattern as the parent clone. It cross-reacts with a major subset of peripheral blood lymphocytes, but not monocytes or granulocytes, of baboon, and rhesus, cynomolgus, and pigtail macaque monkeys. The distribution on lymphocytes is similar to that observed with normal human donor lymphocytes with the majority of CD3-positive cells being negative when dual stained with antibodies to B or NK cells markers. SP34-2 is also capable of inducing cell proliferation on both human and non-human primate PBMC.

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.

560772 Rev. 2
フォーマットの詳細
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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
560772 Rev.2
引用&参考文献
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Development References (8)

  1. Bleavins MR, Brott DA, Alvey JD, de la Iglesia FA. Flow cytometric characterization of lymphocyte subpopulations in the cynomolgus monkey (Macaca fascicularis). Vet Immunol Immunopathol. 1993; 37(1):1-13. (Biology). View Reference
  2. Blumberg RS, Ley S, Sancho J, et al. Structure of the T-cell antigen receptor: evidence for two CD3 epsilon subunits in the T-cell receptor-CD3 complex. Proc Natl Acad Sci U S A. 1990; 87(18):7220-7224. (Clone-specific). View Reference
  3. Conrad ML, Davis WC, Koop BF. TCR and CD3 antibody cross-reactivity in 44 species. Cytometry A. 2007; 71(11):925-933. (Biology). View Reference
  4. Engel P, Wagner N, Tedder TF. CD86 Workshop Report. In: Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995:703-705.
  5. Jacobsen CN, Aasted B, Broe MK, Petersen JL. Reactivities of 20 anti-human monoclonal antibodies with leucocytes from ten different animal species. Vet Immunol Immunopathol. 1993; 39(4):461-466. (Biology). View Reference
  6. Pessano S, Oettgen H, Bhan AK, Terhorst C. The T3/T cell receptor complex: antigenic distinction between the two 20-kd T3 (T3-delta and T3-epsilon) subunits. EMBO J. 1985; 4(2):337-344. (Immunogen). View Reference
  7. Schlossman SF. Stuart F. Schlossman .. et al., ed. Leucocyte typing V : white cell differentiation antigens : proceedings of the fifth international workshop and conference held in Boston, USA, 3-7 November, 1993. Oxford: Oxford University Press; 1995.
  8. Wilson AD, Shooshtari M, Finerty S, Watkins P, Morgan AJ. Selection of monoclonal antibodies for the identification of lymphocyte surface antigens in the New World primate Saguinus oedipus oedipus (cotton top tamarin). J Immunol Methods. 1995; 178(2):195-200. (Biology). View Reference
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560772 Rev. 2

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