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RY610 Rat Anti-Mouse Ly-6C
RY610 Rat Anti-Mouse Ly-6C

Two-color flow cytometric analysis of Ly-6C expression on Mouse splenic leukocytes.  BALB/c Mouse splenocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/32 antibody (Mouse Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with BD Horizon™ BUV395 Rat Anti-Mouse CD8a antibody (Cat. No. 563786) and with either BD Horizon™ RY610 Rat IgM, κ Isotype Control (Cat. No. 571150; Left Plot) or BD Horizon™ RY610 Rat Anti-Mouse Ly-6C antibody (Cat. No. 571148/571149; Right Plot) at 1 μg/test. The bivariate pseudocolor density plot showing the correlated expression of Ly-6C (or Ig Isotype control staining) versus CD8a was derived from gated events with the forward and side light-scatter characteristics of viable splenic leukocytes. Flow cytometry and data analysis were performed using BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.

Two-color flow cytometric analysis of Ly-6C expression on Mouse splenic leukocytes.  BALB/c Mouse splenocytes were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/32 antibody (Mouse Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with BD Horizon™ BUV395 Rat Anti-Mouse CD8a antibody (Cat. No. 563786) and with either BD Horizon™ RY610 Rat IgM, κ Isotype Control (Cat. No. 571150; Left Plot) or BD Horizon™ RY610 Rat Anti-Mouse Ly-6C antibody (Cat. No. 571148/571149; Right Plot) at 1 μg/test. The bivariate pseudocolor density plot showing the correlated expression of Ly-6C (or Ig Isotype control staining) versus CD8a was derived from gated events with the forward and side light-scatter characteristics of viable splenic leukocytes. Flow cytometry and data analysis were performed using BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.

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)
0.2 mg/ml
17067
AB_3686277
Aqueous buffered solution containing ≤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 to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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. An isotype control should be used at the same concentration as the antibody of interest.
  4. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
  5. 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.
  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 http://regdocs.bd.com to access safety data sheets (SDS).
  8. CF™ is a trademark of Biotium, Inc.
  9. For U.S. patents that may apply, see bd.com/patents.
571148 Rev. 1
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.

571148 Rev. 1
Format Details
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RY610
The BD Horizon RealYellow™ 610 (RY610) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 610-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY610 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY610 can be used as an alternative to PE-CF594 and we recommend using an optical filter centered near 610-nm (eg, a 610/20-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE-CF594.
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RY610
Yellow-Green 561 nm
557 nm
610 nm
571148 Rev.1
Citations & References
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View product citations for antibody "571148" on CiteAb

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
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571148 Rev. 1

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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.