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V450 Rat Anti-Mouse Ly-6A/E
V450 Rat Anti-Mouse Ly-6A/E
Flow cytometric analysis of Ly-6A/E on stimulated mouse splenocytes.  BALB/c splenocytes were stimulated with 2.5-5.0 µg/mL concanavilin A (Sigma-Aldrich cat. no. C2010) for 48 hours and were subsequently stained either with a BD Horizon™ V450 Rat IgG2a, κ isotype control (unshaded) or with the BD Horizon™ V450 Rat Anti-Mouse Ly-6A/E antibody (shaded).  Histograms were derived from gated events based on light scattering characteristics for splenocytes.  Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Flow cytometric analysis of Ly-6A/E on stimulated mouse splenocytes.  BALB/c splenocytes were stimulated with 2.5-5.0 µg/mL concanavilin A (Sigma-Aldrich cat. no. C2010) for 48 hours and were subsequently stained either with a BD Horizon™ V450 Rat IgG2a, κ isotype control (unshaded) or with the BD Horizon™ V450 Rat Anti-Mouse Ly-6A/E antibody (shaded).  Histograms were derived from gated events based on light scattering characteristics for splenocytes.  Flow cytometry was performed on a BD™ LSR II flow cytometry system.
Product Details
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BD Horizon™
Ly-6A/E; Lymphocyte antigen 6A-2/6E-1; Ly-6A.2/Ly-6E.1; Sca-1; TAP
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2a, κ
IL-2-dependent mouse T-cell line CTL-L
Flow cytometry (Routinely Tested)
0.2 mg/ml
110454
AB_1727553
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™ V450 under optimum conditions, and unreacted BD Horizon™ V450 was removed.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. BD Horizon V450 has a maximum absorption of 406 nm and maximum emission of 450 nm. Before staining with this reagent, please confirm that your flow cytometer is capable of exciting the fluorochrome and discriminating the resulting fluorescence.
  4. 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.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560653 Rev. 1
Antibody Details
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D7

The D7 monoclonal antibody recognizes Ly-6A.2 and Ly-6E.1, which are allelic members of the Ly-6 multigene family. Sca-1 (Ly6A/E), a phosphatidylinositol-anchored protein of about 18 kDa, is expressed on the multipotent hematopoietic stem cells (HSC) in the bone marrow of mice with both Ly-6 haplotypes. In mice expressing the Ly-6.2 haplotype (e.g., AKR, C57BL, C57BR, C57L, C58, DBA/2, PL, SJL, SWR, 129), Ly-6A/E is also expressed on distinct subpopulations of bone marrow and peripheral B lymphocytes as well as thymic and peripheral T lymphocytes. Strains with the Ly-6.1 haplotype (e.g., A, BALB/c, CBA, C3H/He, DBA/1, NZB) have few Ly-6A/E+ resting peripheral lymphocytes; activation of lymphocytes from mice of both Ly-6 haplotypes leads to strong expression of the Sca-1 antigen. Studies with the D7 antibody have demonstrated that Ly-6A/E may be involved in the regulation of B and T lymphocyte responses, and appears to be required for T-cell receptor-mediated T-cell activation. The purified E13-161.7 mAb (anti-Ly-6A/E) can block binding of FITC-conjugated D7 antibody to mouse splenocytes, but purified mAb D7 is unable to block binding of FITC-conjugated E13-161.7 antibody. Anti-Ly-6A/E (Sca-1) mAb may be used in combination with a Mouse Lineage Panel of antibodies to identify HSC.

The antibody is conjugated to BD Horizon™ V450, 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 Ex max of 406 nm and has an Em Max at 450 nm. Conjugates with BD Horizon™ V450 can be used in place of Pacific Blue™ conjugates.

560653 Rev. 1
Format Details
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V450
BD Horizon™ V450 Dye is part of the BD Horizon™ violet family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 405-nm and an emission maximum (Em Max) at 450-nm. BD Horizon™ V450, driven by BD innovation, is designed to be excited by the violet laser (405 nm) and detected using an optical filter centered near 450-nm (e.g., a 450/50-nm bandpass filter). The dye can be excited by the UV (355-nm) laser resulting in cross-laser excitation and spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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V450
Violet 405 nm
405 nm
450 nm
560653 Rev.1
Citations & References
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Development References (31)

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560653 Rev. 1

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