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RB545 Rat Anti-Mouse Ly-6C
RB545 Rat Anti-Mouse Ly-6C
Two-color flow cytometric analysis of Ly-6C expression on Mouse splenic leucocytes. BALB/c Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse CD8a antibody (Cat. No. 563898) and with either BD Horizon™ RB545 Rat IgM, κ Isotype Control (Cat. No. 569287; Left Plot) or BD Horizon™ RB545 Rat Anti-Mouse Ly-6C (Cat. No. 569288/569289; 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 leucocytes. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 Software. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of Ly-6C expression on Mouse splenic leucocytes. BALB/c Mouse splenocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with BD Horizon™ BV421 Rat Anti-Mouse CD8a antibody (Cat. No. 563898) and with either BD Horizon™ RB545 Rat IgM, κ Isotype Control (Cat. No. 569287; Left Plot) or BD Horizon™ RB545 Rat Anti-Mouse Ly-6C (Cat. No. 569288/569289; 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 leucocytes. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed using FlowJo™ v10.8 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
Aqueous buffered solution containing ≤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 to the dye under optimum conditions and unreacted dye was removed.

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. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  8. For U.S. patents that may apply, see bd.com/patents.
569288 Rev. 2
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.

569288 Rev. 2
Format Details
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RB545
The BD Horizon RealBlue™ 545 (RB545) fluorochrome is part of the BD family of blue (488-nm) laser dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 496-nm and an emission maximum (Em Max) at 545-nm. Driven by BD innovation, RB545 can be used on spectral cytometers and is designed to be excited by the blue (488-nm) laser with minimal excitation by the 561-nm yellow-green laser. RB545 has minimal spillover into yellow-green detectors and a brightness level similar to FITC.  Given its unique emission max, on a spectral instrument RB545 can be used simultaneously with FITC and PE to provide an additional color off of the blue laser. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
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RB545
Blue 488 nm
496 nm
545 nm
569288 Rev.2
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
569288 Rev. 2

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