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BD Pharmingen™ PE-Cy7 Rat Anti-Mouse Ly-6C
Clone HK1.4.rMAb (also known as HK1.4) (RUO)


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Multicolor flow cytometric analysis of Ly-6C expression on Mouse splenic leukocytes. BALB/c Mouse splenic leukocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553142]. The cells were then stained with FITC Rat Anti-Mouse CD8a antibody (Cat. No. 553030; Top Plots) and BD Horizon™ BUV737 Rat Anti-CD11b antibody (Cat. No. 612800; Bottom Plots) and with either PE-Cy7 Rat IgG1, κ Isotype Control (Cat. No. 557645; Left Plots) or PE-Cy7 Rat Anti-Mouse Ly-6C antibody (Cat. No. 569442/569443; Right Plots) at 0.125 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plots showing the correlated expression of Ly-6C (or Ig Isotype control staining) versus CD8a or CD11b were derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) leukocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific
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BD Pharmingen™ PE-Cy7 Rat Anti-Mouse Ly-6C
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Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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 cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- 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.
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- 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).
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
Companion Products






The HK1.4.rMAb monoclonal antibody is a recombinant monoclonal antibody derived from HK1.4 hybridoma cells that specifically recognizes a non-polymorphic determinant on Mouse Lymphocyte antigen Ly-6C which is also known as Ly6c. Ly-6C is an ~14-17 kDa glycosylphosphatidylinositol (GPI)-linked cell-surface antigen that is encoded by Ly6c1 (Lymphocyte antigen 6 complex, locus C1) which belongs to the Ly-6 gene family. Ly-6C is expressed on monocytes, macrophages, neutrophils, eosinophils, endothelial cells, plasma cells, thymocytes, NK cells, and some T cell subsets. Mice with the Ly-6.2 haplotype (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 HK1.4 antibody does not reportedly block the binding of the RB6-8C5 monoclonal antibody that specifically recognizes Mouse Ly-6G and Ly-6C.

Development References (3)
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Havran WL, Lancki DW, Moldwin RL, Dialynas DP, Fitch FW. Characterization of an anti-Ly-6 monoclonal antibody which defines and activates cytolytic T lymphocytes.. J Immunol. 1988; 140(4):1034-42. (Immunogen: Activation, Blocking, (Co)-stimulation, Flow cytometry, Fluorescence activated cell sorting, Functional assay, Stimulation). View Reference
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Ma C, Kapanadze T, Gamrekelashvili J, Manns MP, Korangy F, Greten TF. Anti-Gr-1 antibody depletion fails to eliminate hepatic myeloid-derived suppressor cells in tumor-bearing mice.. J Leukoc Biol. 2012; 92(6):1199-206. (Clone-specific: Flow cytometry). View Reference
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Schlueter AJ, Malek TR, Hostetler CN, Smith PA, deVries P, Waldschmidt TJ. Distribution of Ly-6C on lymphocyte subsets: I. Influence of allotype on T lymphocyte expression.. J Immunol. 1997; 158(9):4211-22. (Biology). View Reference
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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.