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Alexa Fluor® 488 anti-mouse Ly-6G Antibody

Pricing & Availability
Clone
1A8 (See other available formats)
Regulatory Status
RUO
Other Names
Lymphocyte antigen 6 complex, locus G
Isotype
Rat IgG2a, κ
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Product Citations
publications
1A8_A488_Ly-6G_Antibody_1_FC_010516
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.
  • 1A8_A488_Ly-6G_Antibody_1_FC_010516
    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.
  • 1A8_A488_Ly-6G_Antibody_2_120420
    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.
  • 47_Mouse_Lung_Ly6G_CD8_CD31
    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).
See Alexa Fluor® 488 spectral data
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127625 25 µg 124 CHF
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127626 100 µg 265 CHF
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Description

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
Technical Data Sheet (pdf)

Product Details

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

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).

Application References

(PubMed link indicates BioLegend citation)
  1. Fleming TJ, et al. 1993. J. Immunol. 151:2399. (FC)
  2. Daley JM, et al. 2008. J. Leukocyte Biol. 83:1. (FC)
  3. Dietlin TA, et al. 2007. J. Leukocyte Biol. 81:1205. (FC)
  4. Daley J, et al. 2007. J. Leukocyte Biol. doi:10.1189. (Deplete) PubMed
  5. Tadagavadi RK, et al. 2010. J. Immunol. 185:4904. PubMed
  6. Sumagin R, et al. 2010. J. Immunol. 185:7057. PubMed
  7. Guiducci C, et al. 2010. J. Exp Med. 207:2931. PubMed
  8. Fujita M, et al. 2011. Cancer Res. 71:2664. PubMed
  9. Van Leeuwen, et al. 2008. Arterioscler. Thromb. Vasc. Biol. 28:84. (IHC)
  10. Kowanetz M, et al. 2010. P. Natl. Acad. Sci. USA 107:21248. [supplementary data] (IHC)
  11. Esbona K, et al. 2016. Breast Cancer Res. 18:35. (IHC)
  12. Wojtasiak M, et al. 2010. J. Gen. Virol. 91:2158. (FC, Deplete)
  13. Jaeger BN, et al. 2012. J. Exp. Med. 209:565. (Deplete)
  14. Wozniak KL, et al. 2012. BMC Immunol. 13:65 (FC, Deplete)
  15. Ribechini E, et al. 2009. Eur. J. Immunol. 39:3538.
  16. Ng LG, et al. 2011. J Invest. Dermatol. 131:2058. PubMed
  17. Ma C, et al. 2012. J. Leukoc. Biol. 92:1199.
  18. McCartney-Francis, N, et al. 2014. J Leukoc. Biol. 96:917. PubMed
  19. Her Z, et al. 2014. EMBO Mol. Med. 7:24. PubMed
  20. Radtke AJ, et al. 2020. Proc Natl Acad Sci U S A. 117:33455-65. (SB) PubMed
  21. Radtke AJ, et al. 2022. Nat Protoc. 17:378-401. (SB) PubMed
Product Citations
  1. Sweet R, et al. 2017. J Immunol. 10.4049/jimmunol.1600861. PubMed
  2. Bowers E, et al. 2018. Nat Med. 24:95. PubMed
  3. Postat J et al. 2018. Immunity. 49(4):654-665 . PubMed
  4. Pinho–Ribeiro FA, et al. 2018. Cell. 173:1083. PubMed
  5. Groza D, et al. 2018. Oncoimmunology. 7:e1424676. PubMed
  6. Wilson RP, et al. 2019. PLoS Pathog. 15:e1007745. PubMed
  7. Moon H, et al. 2019. Nat Commun. 10:2225. PubMed
  8. Boras M, et al. 2017. J Exp Med. 214:851. PubMed
  9. Sendler M, et al. 2020. Gastroenterology. 158:253. PubMed
  10. Han P, et al. 2020. Sci Adv. 6:eaaz1580. PubMed
  11. Dong X, et al. 2019. ACS Nano. 13:1272. PubMed
  12. Ueki H, et al. 2020. Nat Protoc. 15:1041. PubMed
  13. Joy MT, et al. 2019. Cell. 176:1143. PubMed
  14. Liu S, et al. 2020. Neuron. S0896-6273(20)30532-8.. PubMed
  15. Georgoudaki A, et al. 2016. Cell Rep. 15: 2000-2011. PubMed
  16. Green D, et al. 2020. Oncogene. 39:5553. PubMed
  17. Dudeck J, et al. 2021. Immunity. 54(3):468-483.e5. PubMed
  18. Barkaway A, et al. 2021. Immunity. . PubMed
  19. Souza COS, et al. 2021. iScience. 24(6):102548. PubMed
  20. Du J, et al. 2021. Mol Med Rep. 23:. PubMed
  21. Pach E, et al. 2021. Cancers (Basel). 13:. PubMed
  22. Myerson JW, et al. 2021. Nat Nanotechnol. Online ahead of print. PubMed
  23. Craver BM, et al. 2020. Blood Adv. 4:312. PubMed
  24. Garland KM, et al. 2021. Front Immunol. 12:753472. PubMed
  25. Almutairi F, et al. 2021. Front Immunol. 12:772288. PubMed
  26. Wu L, et al. 2022. Theranostics. 12:842. PubMed
  27. Stolp B, et al. 2022. Cell Rep. 38:110387. PubMed
  28. Liu G, et al. 2022. Ann Transl Med. 10:693. PubMed
  29. Takizawa S, et al. 2022. Mol Med. 28:91. PubMed
  30. Jahan S, et al. 2022. J Biol Chem. :102386. PubMed
  31. Leung H, et al. 2022. Nat Commun. 13:5206. PubMed
  32. El-Naccache DW, et al. 2022. Cell Rep. 40:111150. PubMed
  33. Abdelnabi MN, et al. 2022. Cell Mol Gastroenterol Hepatol. 14:1269. PubMed
  34. Lee DE, et al. 2022. Nat Commun. 13:7613. PubMed
  35. Ullah I, et al. 2023. Cell Rep Med. 4:100893. PubMed
  36. Cagle LA, et al. 2023. Front Physiol. 13:1059686. PubMed
  37. Gershater M, et al. 2022. J Immunol. 208:1595. PubMed
  38. Gupta T, et al. 2023. Front Immunol. 14:1044703. PubMed
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
UniProt
View information about Ly-6G on UniProt.org

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?

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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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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