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BV421 Rat Anti-Mouse Ly-6C
BV421 Rat Anti-Mouse Ly-6C
Multicolor flow cytometric analysis of Ly-6C expression on mouse splenocytes.  Splenocytes from a BALB/c mouse were stained with FITC Rat Anti-Mouse CD8a antibody (Cat. No. 553031/553030/561966) and with either BD Horizon™ BV421 Rat IgM, κ Isotype Control (Cat. No. 562708, Left Panel) or with BD Horizon™ BV421 Rat Anti-Mouse Ly-6C antibody (Cat. No. 562727, Right Panel). Two-color flow cytometric dot plots showing the correlated expression of Ly-6C (or Ig isotype control staining) versus CD8a were derived from gated events with the forward and side light-scattering characteristics of viable splenocytes. Flow cytometry was performed with a BD™ LSR II Flow Cytometry System.
BV421 Rat Anti-Mouse Ly-6C
Immunohistofluorescent analysis of Ly-6C expression by cells within C57BL/6 mouse spleen. A mouse spleen cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with BD Horizon™ BV421 Rat Anti-Mouse Ly-6C antibody (Cat. No. 562727, pseudo-colored green) and Alexa Fluor® 488 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557669, pseudo-colored red). Images were captured on a standard epifluorescence microscope. Original magnification, 20x.
Multicolor flow cytometric analysis of Ly-6C expression on mouse splenocytes.  Splenocytes from a BALB/c mouse were stained with FITC Rat Anti-Mouse CD8a antibody (Cat. No. 553031/553030/561966) and with either BD Horizon™ BV421 Rat IgM, κ Isotype Control (Cat. No. 562708, Left Panel) or with BD Horizon™ BV421 Rat Anti-Mouse Ly-6C antibody (Cat. No. 562727, Right Panel). Two-color flow cytometric dot plots showing the correlated expression of Ly-6C (or Ig isotype control staining) versus CD8a were derived from gated events with the forward and side light-scattering characteristics of viable splenocytes. Flow cytometry was performed with a BD™ LSR II Flow Cytometry System.
Immunohistofluorescent analysis of Ly-6C expression by cells within C57BL/6 mouse spleen. A mouse spleen cryosection (5 µm) was fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), blocked with 5% goat serum and 1% BSA diluted in 1x PBS, and stained with BD Horizon™ BV421 Rat Anti-Mouse Ly-6C antibody (Cat. No. 562727, pseudo-colored green) and Alexa Fluor® 488 Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 557669, pseudo-colored red). Images were captured on a standard epifluorescence microscope. Original magnification, 20x.
Product Details
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BD Horizon™
Ly6c; Lymphocyte antigen 6 complex, locus C; Lymphocyte antigen Ly-6C
Mouse (QC Testing)
Rat IgM, κ
Not reported
Flow cytometry (Routinely Tested), Immunofluorescence (Tested During Development)
0.2 mg/ml
AB_2737748
Aqueous buffered solution containing BSA 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 BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
562727 Rev. 3
Antibody Details
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AL-21

The AL-21 monoclonal antibody specifically binds to a non-polymorphic determinant of Ly-6C, a 14-17 kDa GPI-linked cell-surface antigen found on some monocyte/macrophage populations, granulocytes, endothelial cells, plasma cells, and thymocyte, NK-cell, and T-subsets.  Mice with the Ly-6.2 alloantigen (eg, AKR, C57BL, C57BR, C57L, C58, DBA/2, PL, SJL, SWR, 129) have subsets of CD8+ and CD4+ Ly-6C+ T cells, while Ly-6.1 strains (eg, A, BALB/c, CBA, C3H/He, DBA/1, NZB) have only CD8+ Ly-6C+ T cells.   Upregulation of Ly-6C expression on CD8+ T cells by interferons α and β and poly (I:C) has been described,  and Ly-6C is a memory marker on CD8+ T cells.

The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon™ BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon™ BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates.

562727 Rev. 3
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
562727 Rev.3
Citations & References
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Development References (7)

  1. Cerwenka A, Carter LL, Reome JB, Swain SL, Dutton RW. In vivo persistence of CD8 polarized T cell subsets producing type 1 or type 2 cytokines. J Immunol. 1998; 161(1):97-105. (Biology). View Reference
  2. Jutila DB, Kurk S, Jutila MA.. Differences in the expression of Ly-6C on neutrophils and monocytes following PI-PLC hydrolysis and cellular activation.. Immunol Lett. 1994 ; 41(1):49-57. (Biology). View Reference
  3. Jutila MA, Kroese FG, Jutila KL, et al. Ly-6C is a monocyte/macrophage and endothelial cell differentiation antigen regulated by interferon-gamma. Eur J Immunol. 1988; 18(11):1819-1826. (Biology). View Reference
  4. Sato N, Yahata T, Santa K. Functional characterization of NK1.1 + Ly-6C+ cells. Immunol Lett. 1996; 1(54):1-5-9. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  5. Takahama Y, Sharrow SO, Singer A. Expression of an unusual T cell receptor (TCR)-V beta repertoire by Ly-6C+ subpopulations of CD4+ and/or CD8+ thymocytes. Evidence for a developmental relationship between Ly-6C+ thymocytes and CD4-CD8-TCR-alpha beta+ thymocytes. J Immunol. 1991; 147(9):2883-2891. (Clone-specific: Flow cytometry). View Reference
  6. Tough DF, Borrow P, Sprent J. Induction of bystander T cell proliferation by viruses and type I interferon in vivo. Science. 1996; 272(5270):1947-1950. (Biology). View Reference
  7. Wrammert J, Källberg E, Agace WW, Leanderson T. Ly6C expression differentiates plasma cells from other B cell subsets in mice. Eur J Immunol. 2002; 32(1):97-103. (Clone-specific: Flow cytometry). View Reference
View All (7) View Less
562727 Rev. 3

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