- 1A8 (See other available formats)
- Regulatory Status
- Other Names
- Lymphocyte antigen 6 complex, locus G
- Rat IgG2a, κ
- Ave. Rating
- Submit a Review
- Product Citations
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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.Product Details
- Antibody Type
- Host Species
- Ly-6G transfected EL-4J cell line.
- Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
- The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 488 under optimal conditions.
- 0.5 mg/mL
- Storage & Handling
- The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
FC - Quality tested
IHC-F - Verified
SB - Reported in the literature, not verified in house
- Recommended Usage
Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is ≤ 0.5 µg per million cells in 100 µL volume. For immunohistochemistry on frozen tissue sections, a concentration range of 2.5 - 10 μg/mL is recommended. It is recommended that the reagent be titrated for optimal performance for each application.
* Alexa Fluor® 488 has a maximum emission of 519 nm when it is excited at 488 nm.
Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.
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- Excitation Laser
Blue Laser (488 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).
(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
- Her Z, et al. 2014. EMBO Mol. Med. 7:24. PubMed
- Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
- Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
- Product Citations
AB_2561339 (BioLegend Cat. No. 127625)
AB_2561340 (BioLegend Cat. No. 127626)
- A 21-35 kD GPI-anchorded membrane protein
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.
- Cell Type
- Granulocytes, Macrophages, Monocytes
- Biology Area
- Immunology, Innate Immunity
- Antigen References
Fleming TJ, et al. 1993. J. Immunol. 151:2399.
- Gene ID
- 546644 View all products for this Gene ID
- View information about Ly-6G on UniProt.org
- If an antibody clone has been previously successfully used in IBEX in one fluorescent format, will other antibody formats work as well?
It’s likely that other fluorophore conjugates to the same antibody clone will also be compatible with IBEX using the same sample fixation procedure. Ultimately a directly conjugated antibody’s utility in fluorescent imaging and IBEX may be specific to the sample and microscope being used in the experiment. Some antibody clone conjugates may perform better than others due to performance differences in non-specific binding, fluorophore brightness, and other biochemical properties unique to that conjugate.
- Will antibodies my lab is already using for fluorescent or chromogenic IHC work in IBEX?
Fundamentally, IBEX as a technique that works much in the same way as single antibody panels or single marker IF/IHC. If you’re already successfully using an antibody clone on a sample of interest, it is likely that clone will have utility in IBEX. It is expected some optimization and testing of different antibody fluorophore conjugates will be required to find a suitable format; however, legacy microscopy techniques like chromogenic IHC on fixed or frozen tissue is an excellent place to start looking for useful antibodies.
- Are other fluorophores compatible with IBEX?
Over 18 fluorescent formats have been screened for use in IBEX, however, it is likely that other fluorophores are able to be rapidly bleached in IBEX. If a fluorophore format is already suitable for your imaging platform it can be tested for compatibility in IBEX.
- The same antibody works in one tissue type but not another. What is happening?
Differences in tissue properties may impact both the ability of an antibody to bind its target specifically and impact the ability of a specific fluorophore conjugate to overcome the background fluorescent signal in a given tissue. Secondary stains, as well as testing multiple fluorescent conjugates of the same clone, may help to troubleshoot challenging targets or tissues. Using a reference control tissue may also give confidence in the specificity of your staining.
- How can I be sure the staining I’m seeing in my tissue is real?
In general, best practices for validating an antibody in traditional chromogenic or fluorescent IHC are applicable to IBEX. Please reference the Nature Methods review on antibody based multiplexed imaging for resources on validating antibodies for IBEX.
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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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Alexa Fluor® 594 anti-mouse Ly-6G
Purified anti-mouse Ly-6G
Biotin anti-mouse Ly-6G
FITC anti-mouse Ly-6G
PE anti-mouse Ly-6G
Alexa Fluor® 647 anti-mouse Ly-6G
Pacific Blue™ anti-mouse Ly-6G
APC anti-mouse Ly-6G
PerCP/Cyanine5.5 anti-mouse Ly-6G
PE/Cyanine7 anti-mouse Ly-6G
Alexa Fluor® 700 anti-mouse Ly-6G
APC/Cyanine7 anti-mouse Ly-6G
Alexa Fluor® 488 anti-mouse Ly-6G
Brilliant Violet 421™ anti-mouse Ly-6G
Brilliant Violet 570™ anti-mouse Ly-6G
Ultra-LEAF™ Purified anti-mouse Ly-6G
Brilliant Violet 510™ anti-mouse Ly-6G
Purified anti-mouse Ly-6G (Maxpar® Ready)
Brilliant Violet 650™ anti-mouse Ly-6G
Brilliant Violet 711™ anti-mouse Ly-6G
Brilliant Violet 605™ anti-mouse Ly-6G
Brilliant Violet 785™ anti-mouse Ly-6G
PE/Dazzle™ 594 anti-mouse Ly-6G
APC/Fire™ 750 anti-mouse Ly-6G
PerCP anti-mouse Ly-6G
TotalSeq™-A0015 anti-mouse Ly-6G
TotalSeq™-C0015 anti-mouse Ly-6G
TotalSeq™-B0015 anti-mouse Ly-6G
Spark Blue™ 550 anti-mouse Ly-6G
Spark NIR™ 685 anti-mouse Ly-6G
Spark YG™ 593 anti-mouse Ly-6G
PE/Cyanine5 anti-mouse Ly-6G
PE/Fire™ 810 anti-mouse Ly-6G Antibody