Flow cytometric analysis of Ly-6C on mouse splenocytes. Splenocytes from BALB/c mice were stained either with a APC-Cy™7 Rat IgM, κ isotype control (Cat. No. 560571, left panel) or with the APC-Cy™7 Rat Anti-Mouse Ly-6C antibody (Cat. No. 560596, right panel) in conjunction with a FITC Rat Anti-Mouse CD8a antibody (Cat. No. 553031). Dot plots 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-6C on mouse splenocytes. Splenocytes from BALB/c mice were stained either with a APC-Cy™7 Rat IgM, κ isotype control (Cat. No. 560571, left panel) or with the APC-Cy™7 Rat Anti-Mouse Ly-6C antibody (Cat. No. 560596, right panel) in conjunction with a FITC Rat Anti-Mouse CD8a antibody (Cat. No. 553031). Dot plots 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
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_1727555
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 APC-Cy7 under optimum conditions, and unconjugated antibody and free APC-Cy7 were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- 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).
- 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.
- APC-Cy7 tandem fluorochrome emission is collected in a detector for fluorescence wavelengths of 750 nm and higher.
- APC-Cy7 is a tandem fluorochrome composed of Allophycocyanin (APC), which is excited by laser lines between 595 and 647 nm and serves as an energy donor, coupled to the cyanine dye Cy7™, which acts as an energy acceptor and fluoresces at 780 nm. BD Biosciences Pharmingen has maximized the fluorochrome energy transfer in APC-Cy7, thus maximizing its fluorescence emission intensity, minimizing residual emission from APC, and minimizing required electronic compensation in multilaser-laser flow cytometry systems. Note: Although every effort is made to minimize the lot-to-lot variation in residual emission from APC, it is strongly recommended that every lot be tested for differences in the amount of compensation required and that individual compensation controls are run for each APC-Cy7 conjugate.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Cy is a trademark of GE Healthcare.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
APC-Cy™7 Rat IgM, κ Isotype Control RUO
Size
0.1 mg
Cat No.
560571
FITC Rat Anti-Mouse CD8a RUO
Size
0.5 mg
Cat No.
553031
560596 Rev. 2
Antibody Details
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.
560596 Rev. 2
Format Details
APC-Cy7
APC-Cy7 dye is a part of the BD APC red family of dyes. This tandem fluorochrome is comprised of a Allophycocyanin (APC) donor that has excitation maxima (Ex Max) of 651 nm and an acceptor dye, Cy™7, with an emission maximum (Em Max) at 779 nm. APC-Cy7 can be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 780 nm (e.g., a 760/60 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
APC-Cy7
Red 627-640 nm
651 nm
779 nm
560596 Rev.2
Citations & References
Development References (7)
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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
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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
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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
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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
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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
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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
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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
560596 Rev. 2
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.
