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BD Horizon™ V500 Syrian Hamster anti-Mouse CD3e
Clone 500A2 (RUO)
CD3e expression in spleen. Splenocytes from BALB/c mice were simultaneously stained with PE rat anti-mouse CD4 mAb RM4-5 (Cat. no. 553048), PE rat anti-mouse CD8a mAb 53-6.7 (Cat. no. 553033), and Horizon™ V500 Hamster IgG2, λ Isotype Control (Cat. no. 560785, left panel) or Horizon™ V500 Hamster anti-Mouse CD3e (right panel) in the presence of BD Fc Block™ (Cat. no. 553142). Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
CD3e expression in spleen. Splenocytes from BALB/c mice were simultaneously stained with PE rat anti-mouse CD4 mAb RM4-5 (Cat. no. 553048), PE rat anti-mouse CD8a mAb 53-6.7 (Cat. no. 553033), and Horizon™ V500 Hamster IgG2, λ Isotype Control (Cat. no. 560785, left panel) or Horizon™ V500 Hamster anti-Mouse CD3e (right panel) in the presence of BD Fc Block™ (Cat. no. 553142). Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
CD3e expression in spleen. Splenocytes from BALB/c mice were simultaneously stained with PE rat anti-mouse CD4 mAb RM4-5 (Cat. no. 553048), PE rat anti-mouse CD8a mAb 53-6.7 (Cat. no. 553033), and Horizon™ V500 Hamster IgG2, λ Isotype Control (Cat. no. 560785, left panel) or Horizon™ V500 Hamster anti-Mouse CD3e (right panel) in the presence of BD Fc Block™ (Cat. no. 553142). Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
CD3e expression in spleen. Splenocytes from BALB/c mice were simultaneously stained with PE rat anti-mouse CD4 mAb RM4-5 (Cat. no. 553048), PE rat anti-mouse CD8a mAb 53-6.7 (Cat. no. 553033), and Horizon™ V500 Hamster IgG2, λ Isotype Control (Cat. no. 560785, left panel) or Horizon™ V500 Hamster anti-Mouse CD3e (right panel) in the presence of BD Fc Block™ (Cat. no. 553142). Flow cytometry was performed on a BD FACSCanto™ II flow cytometry system.
Product Details
BD Horizon™
CD3; CD3 epsilon; Cd3e; CD3ε; T3e
Mouse (QC Testing)
Syrian Hamster IgG2, κ
Mouse T-cell receptor
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_1937314
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
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Companion Products
Purified Rat Anti-Mouse CD16/CD32 (Mouse BD Fc Block™) RUO
Size
0.5 mg
Cat No.
553142
PE Rat Anti-Mouse CD4 RUO
Size
0.1 mg
Cat No.
553048
PE Rat Anti-Mouse CD8a RUO
Size
0.2 mg
Cat No.
553033
560773 Rev. 1
Antibody Details
500A2
The 500A2 monoclonal antibody specifically binds to the 25-kDa ε chain of the T-cell receptor-associated CD3 complex expressed on mouse thymocytes, mature T lymphocytes, and NKT cells. Plate-bound and soluble forms of the 500A2 antibody can activate T cells in vitro. Activation of a mouse T-cell clone by the 500A2 antibody can be blocked by Fab fragments of the GK1.5 anti-CD4 antibody. This suggests that the 500A2 antibody may bind an epitope on CD3e close to a site at which CD4 associates with the T-cell receptor. The 500A2 antibody reportedly does not to crossreact with rat leukocytes.
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.
560773 Rev. 1
Format Details
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.When compensating dyes in this spectral range (such as BD 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 (eg, 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.
V500
415 nm
499 nm
560773 Rev.1
Citations & References
View product citations for antibody "560773" on CiteAb
Development References (5)
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Allison JP, Havran WL, Poenie M, et al. Expression and function of CD3 on murine thymocytes. In: Kappler J, Davis M, ed. The T-Cell Receptor, UCLA Symposia, 73rd Edition. Los Angeles: 1988:33-45.
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Havran WL, Poenie M, Kimura J, Tsien R, Weiss A, Allison JP. Expression and function of the CD3-antigen receptor on murine CD4+8+ thymocytes. Nature. 1987; 330(6144):170-173. (Clone-specific). View Reference
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Kubo RT, Born W, Kappler JW, Marrack P, Pigeon M. Characterization of a monoclonal antibody which detects all murine alpha beta T cell receptors. J Immunol. 1989; 142(8):2736-2742. (Methodology: Flow cytometry). View Reference
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Ortaldo JR, Winkler-Pickett R, Mason AT, Mason LH. The Ly-49 family: regulation of cytotoxicity and cytokine production in murine CD3+ cells. J Immunol. 1998; 160(1):1158-1165. (Clone-specific). View Reference
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Portoles P, Rojo J, Golby A, et al . Monoclonal antibodies to murine CD3 epsilon define distinct epitopes, one of which may interact with CD4 during T cell activation. J Immunol. 1989; 142(12):4169-4175. (Clone-specific). View Reference
560773 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.
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.