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PE-Cy™7 Rat Anti-Mouse Ly-6C
PE-Cy™7 Rat Anti-Mouse Ly-6C

Flow cytometric analysis of Ly-6C on mouse splenocytes. BALB/c mice Splenocytes were stained either with a PE-Cy™7 Rat IgM, κ Isotype Control (Cat. No. 560572, left panel) or with the PE-Cy™7 Rat Anti-Mouse Ly-6C (Cat. No. 560593, right panel) in conjunction with a FITC Rat Anti-Mouse CD8a (Cat. No.553031/561966/553030). The fluorescence dot plots were derived from gated events with the light scattering characteristics of viable splenocytes. Flow cytometry was performed on a BD™ LSR II.

Flow cytometric analysis of Ly-6C on mouse splenocytes. BALB/c mice Splenocytes were stained either with a PE-Cy™7 Rat IgM, κ Isotype Control (Cat. No. 560572, left panel) or with the PE-Cy™7 Rat Anti-Mouse Ly-6C (Cat. No. 560593, right panel) in conjunction with a FITC Rat Anti-Mouse CD8a (Cat. No.553031/561966/553030). The fluorescence dot plots were derived from gated events with the light scattering characteristics of viable splenocytes. Flow cytometry was performed on a BD™ LSR II.

Product Details
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BD Pharmingen™
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
AB_1727557
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 PE-Cy7 under optimum conditions, and unconjugated antibody and free PE-Cy7 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. 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. Cy is a trademark of GE Healthcare.
  5. Warning: Some APC-Cy7 and PE-Cy7 conjugates show changes in their emission spectrum with prolonged exposure to formaldehyde. If you are unable to analyze fixed samples within four hours, we recommend that you use BD™ Stabilizing Fixative (Cat. No. 338036).
  6. 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.
  7. PE-Cy7 is a tandem fluorochrome composed of R-phycoerythrin (PE), which is excited by 488-nm light and serves as an energy donor, coupled to the cyanine dye Cy7, which acts as an energy acceptor and fluoresces maximally at 780 nm. PE-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from PE may be observed. Therefore, we recommend that individual compensation controls be performed for every PE-Cy7 conjugate. PE-Cy7 is optimized for use with a single argon ion laser emitting 488-nm light, and there is no significant overlap between PE-Cy7 and FITC emission spectra. When using dual-laser cytometers, which may directly excite both PE and Cy7, we recommend the use of cross-beam compensation during data acquisition or software compensation during data analysis.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
560593 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.

560593 Rev. 2
Format Details
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PE-Cy7
PE-Cy7 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 781-nm. PE can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and detected using an optical filter centered near 781 nm (e.g., a 760/60-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the Red (627–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE-Cy7
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
781 nm
560593 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. (Biology). 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. (Biology). 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. (Biology). View Reference
View All (7) View Less
560593 Rev. 2

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