C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS for 1 hour at room temperature. Then the section was stained with 5 µg/mL of anti-mouse Ly-6G (clone 1A8) Spark YG™ 570 (red), 5 µg/mL of anti-mouse CDε Alexa Fluor® 647 (green) overnight at 4°C. The nuclei were counterstained with DAPI (blue). The image was captured by 10X objective.
C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS for 1 hour at room temperature. Then the section was stained with 5 µg/mL of anti-mouse Ly-6G (clone 1A8) Spark YG™ 570 (red), 5 µg/mL of anti-mouse CDε Alexa Fluor® 647 (green) overnight at 4°C. The nuclei were counterstained with DAPI (blue). The image was captured by 10X objective.
C57BL/6 mouse frozen cerebellum section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS for 1 hour at room temperature. Then the section was stained with 5 µg/mL of anti-mouse Ly-6G (clone 1A8) Spark YG™ 570 (negative) overnight at 4°C. The nuclei were counterstained with DAPI (blue). The image was captured by 10X objective.
C57BL/6 mouse bone marrow cells were stained with anti-mouse Ly-6G (clone 1A8) Spark YG™ 570 (filled histogram) or rat IgG2a, κ Spark YG™ 570 isotype control (open histogram). Data shown was gated on myeloid cell population.
Lymphocyte antigen 6 complex, locus G (Ly-6G), a 21-25 kD GPI-anchored protein, is expressed on the majority of myeloid cells in bone marrow and peripheral granulocytes.
For immunohistochemistry on frozen tissue sections, a concentration range of 5.0 - 10.0 µg/mL is suggested. For flow cytometric staining, the suggested use of this reagent is ≤ 0.25 µg per million cells in 100 µL volume. It is recommended that the reagent be titrated for optimal performance for each application.
* Spark YG™ 570 has a maximum excitation of 555 nm and a maximum emission of 570 nm.
Excitation Laser
Green Laser (532 nm)/Yellow-Green Laser (561 nm)
Application Notes
While 1A8 recognizes only Ly-6G, clone RB6-8C5 recognizes both Ly-6G and Ly-6C. Clone RB6-8C5 binds with high affinity to mouse Ly-6G molecules and to a lower extent to Ly-6C15. Clone RB6-8C5 impairs the binding of anti-mouse Ly-6G clone 1A815. However, clone RB6-8C5 is able to stain in the presence of anti-mouse Ly-6C clone HK1.416.
Additional reported applications (for the relevant formats) include: immunohistochemistry9 of frozen sections10 and paraffin-embedded sections11, depletion4, 12-14, and spatial biology (IBEX)20,21. The Ultra-LEAF™ purified antibody (Endotoxin < 0.01 EU/µg, Azide-Free, 0.2 µm filtered) is recommended for in vivo studies or highly sensitive assays (Cat. No. 127632, 127649, 127650, 127661 and 127662).
Application References
(PubMed link indicates BioLegend citation)
Fleming TJ, et al. 1993. J. Immunol. 151:2399. (FC)
Daley JM, et al. 2008. J. Leukocyte Biol. 83:1. (FC)
Dietlin TA, et al. 2007. J. Leukocyte Biol. 81:1205. (FC)
Daley J, et al. 2007. J. Leukocyte Biol. doi:10.1189. (Deplete) PubMed
Tadagavadi RK, et al. 2010. J. Immunol. 185:4904. PubMed
Sumagin R, et al. 2010. J. Immunol. 185:7057. PubMed
Guiducci C, et al. 2010. J. Exp Med. 207:2931. PubMed
Fujita M, et al. 2011. Cancer Res. 71:2664. PubMed
Van Leeuwen, et al. 2008. Arterioscler. Thromb. Vasc. Biol. 28:84. (IHC)
Kowanetz M, et al. 2010. P. Natl. Acad. Sci. USA 107:21248. [supplementary data] (IHC)
Esbona K, et al. 2016. Breast Cancer Res. 18:35. (IHC)
Wojtasiak M, et al. 2010. J. Gen. Virol. 91:2158. (FC, Deplete)
Jaeger BN, et al. 2012. J. Exp. Med. 209:565. (Deplete)
Wozniak KL, et al. 2012. BMC Immunol. 13:65 (FC, Deplete)
Ribechini E, et al. 2009. Eur. J. Immunol. 39:3538.
Ng LG, et al. 2011. J Invest. Dermatol. 131:2058. PubMed
Ma C, et al. 2012. J. Leukoc. Biol. 92:1199.
McCartney-Francis, N, et al. 2014. J Leukoc. Biol. 96:917. PubMed
Expressed on the majority of myeloid cells in bone marrow and peripheral granulocytes. The monoclonal antibody RB6-8C5 recognizes both Ly-6G and Ly-6C.
This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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