- Clone
- 1A8 (See other available formats)
- Regulatory Status
- RUO
- Other Names
- Lymphocyte antigen 6 complex, locus G
- Isotype
- Rat IgG2a, κ
- Ave. Rating
- Submit a Review
- Product Citations
- publications
![1A8_A488_Ly-6G_Antibody_1_FC_010516 1A8_A488_Ly-6G_Antibody_1_FC_010516](https://d1spbj2x7qk4bg.cloudfront.net/Admin/Public/GetImage.ashx?Image=/Files/Images/media_assets/products/product_images/1A8_A488_Ly-6G_Antibody_1_FC_010516.jpg&Width=240&Height=300&altFmImage_path=&Compression=90&Crop=5)
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C57BL/6 mouse bone marrow cells were stained with Ly-6G (clone 1A8) Alexa Fluor® 488 (filled histogram) or rat IgG2a, κ Alexa Fluor® 488 isotype control (open histogram). Data shown was gated on the myeloid cell population. -
C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for ten minutes at room temperature and blocked with 5% FBS plus 5% rat/mouse serum for one hour at room temperature. Then the section was stained with 2.5 µg/ml of Ly-6G (clone 1A8) Alexa Fluor® 488 (green), 5 µg/ml of CD3 (clone 17A2) Alexa Fluor® 647 (red), and 5 µg/ml of B220 (clone RA3-6B2) Brilliant Violet 421™ (blue) overnight at 4°C. The image was captured by 10X objective. -
Confocal image of C57BL/6 mouse lung sample acquired using the IBEX method of highly multiplexed antibody-based imaging: Ly-6G (yellow) in Cycle 2, CD31 (blue) in Cycle 3, and CD8 (red) in Cycle 4. Tissues were prepared using ~1% (vol/vol) formaldehyde and a detergent. Following fixation, samples are immersed in 30% (wt/vol) sucrose for cryoprotection. Images are courtesy of Drs. Andrea J. Radtke and Ronald N. Germain of the Center for Advanced Tissue Imaging (CAT-I) in the National Institute of Allergy and Infectious Diseases (NIAID, NIH).
Cat # | Size | Price | Quantity Check Availability | Save | ||
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127625 | 25 µg | 124 CHF | ||||
127626 | 100 µg | 265 CHF |
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 DetailsProduct Details
- Verified Reactivity
- Mouse
- Antibody Type
- Monoclonal
- Host Species
- Rat
- Immunogen
- Ly-6G transfected EL-4J cell line.
- Formulation
- Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
- Preparation
- The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 488 under optimal conditions.
- Concentration
- 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.
- Application
-
FC - Quality tested
IHC-F - Verified
SB - Reported in the literature, not verified in house - Recommended Usage
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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.
View full statement regarding label licenses - 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). -
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
- 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
-
- RRID
-
AB_2561339 (BioLegend Cat. No. 127625)
AB_2561340 (BioLegend Cat. No. 127626)
Antigen Details
- Structure
- A 21-35 kD GPI-anchorded membrane protein
- Distribution
-
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
- Specificity (DOES NOT SHOW ON TDS):
- Ly-6G
- Specificity Alt (DOES NOT SHOW ON TDS):
- Ly-6G
- App Abbreviation (DOES NOT SHOW ON TDS):
- FC,IHC-F,SB
- UniProt
- View information about Ly-6G on UniProt.org
Related Pages & Pathways
Pages
Related FAQs
- 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?
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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.
Other Formats
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Compare Data Across All Formats
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.
If you need assistance with selecting the best format contact our expert technical support team.
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Alexa Fluor® 594 anti-mouse Ly-6G
C57BL/6 mouse frozen spleen section was fixed with 4% parafo... -
Purified anti-mouse Ly-6G
C57BL/6 bone marrow cells stained with 1A8 purified, followe... Fresh, frozen mouse spleen was stained with purified Ly6G cl... Mouse uterine tissue fixed in 10% formalin, paraffin embedde... -
Biotin anti-mouse Ly-6G
C57BL/6 bone marrow cells stained with 1A8 biotin, followed ... -
FITC anti-mouse Ly-6G
BALB/cmouse bone marrow cells stained with 1A8 FITC (gated o... -
PE anti-mouse Ly-6G
C57BL/6 bone marrow cells stained with 1A8 PE (gated on myel... -
Alexa Fluor® 647 anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells stained with 1A8 Alexa Fluor... C57BL/6 mouse frozen spleen section was fixed with 4% parafo... -
Pacific Blue™ anti-mouse Ly-6G
C57BL/6 bone marrow cells stained with 1A8 Pacific Blue&trad... -
APC anti-mouse Ly-6G
C57BL/6 bone marrow cells stained with 1A8 APC -
PerCP/Cyanine5.5 anti-mouse Ly-6G
C57BL/6 bone marrow cells stained with 1A8 PerCP/Cyanine5.5 -
PE/Cyanine7 anti-mouse Ly-6G
C57BL/6 bone marrow cells stained with 1A8 PE/Cyanine7 -
Alexa Fluor® 700 anti-mouse Ly-6G
C57BL/6 bone marrow cells stained with 1A8 Alexa Fluor® ... -
APC/Cyanine7 anti-mouse Ly-6G
C57BL/6 bone marrow cells stained with 1A8 APC/Cyanine7 -
Alexa Fluor® 488 anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with Ly-6G (clo... C57BL/6 mouse frozen spleen section was fixed with 4% parafo... Confocal image of C57BL/6 mouse lung sample acquired using t... -
Brilliant Violet 421™ anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with Ly-6G (clo... C57BL/6 mouse frozen spleen section was fixed with 4% parafo... -
Brilliant Violet 570™ anti-mouse Ly-6G
C57BL/6 bone marrow cells were stained with Ly-6G (clone 1A8... -
Ultra-LEAF™ Purified anti-mouse Ly-6G
C57BL/6 bone marrow cells stained with 1A8 purified, followe... Mouse uterine tissue fixed in 10% formalin, paraffin embedde... -
Brilliant Violet 510™ anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with Ly-6G (clo... -
Purified anti-mouse Ly-6G (Maxpar® Ready)
C57BL/6 mouse bone marrow cells stained with 148Nd-anti-CD11... Western blot analysis of mouse bone marrow (lane 1), mouse s... -
Brilliant Violet 650™ anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with Ly-6G (clo... -
Brilliant Violet 711™ anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with Ly-6G (clo... -
Brilliant Violet 605™ anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with Ly-6G (clo... -
Brilliant Violet 785™ anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with Ly-6G (clo... -
PE/Dazzle™ 594 anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with Ly-6G (clo... -
APC/Fire™ 750 anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with Ly-6G (clo... -
PerCP anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with Ly-6G (clo... -
TotalSeq™-A0015 anti-mouse Ly-6G
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TotalSeq™-C0015 anti-mouse Ly-6G
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TotalSeq™-B0015 anti-mouse Ly-6G
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Spark Blue™ 550 anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells stained with Ly-6G (clone 1A... -
Spark NIR™ 685 anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with Ly-6G (clo... -
Spark YG™ 593 anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with anti-mouse... -
APC/Fire™ 810 anti-mouse Ly-6G Antibody
C57BL/6 mouse bone marrow cells were stained with anti-mouse... -
PE/Cyanine5 anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with anti-mouse... -
PE/Fire™ 810 anti-mouse Ly-6G Antibody
C57BL/6 mouse bone marrow cells were stained with anti-mouse... -
Spark UV™ 387 anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with anti-mouse... -
PE/Fire™ 640 anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells stained with anti-mouse Ly-6... -
Spark YG™ 570 anti-mouse Ly-6G
C57BL/6 mouse bone marrow cells were stained with anti-mouse... C57BL/6 mouse frozen spleen section was fixed with 4% parafo... C57BL/6 mouse frozen cerebellum section was fixed with 4% pa...
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