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Alexa Fluor® 594 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
IA8_A594_Ly-6G_Antibody_1_112520
C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS plus 5% rat serum for 1 hour at room temperature. Then the section was stained with 2.5 µg/ml of Ly-6G (clone 1A8) Alexa Fluor® 594 (red), 2.5 µg/ml of CD3 (clone 145-2C11) Alexa Fluor® 647 (blue), and 2.5 µg/ml of B220 (clone RA3-6B2) Alexa Fluor® 488 (green) overnight at 4°C. The image was captured by 10X objective.
  • IA8_A594_Ly-6G_Antibody_1_112520
    C57BL/6 mouse frozen spleen section was fixed with 4% paraformaldehyde (PFA) for 10 minutes at room temperature and blocked with 5% FBS plus 5% rat serum for 1 hour at room temperature. Then the section was stained with 2.5 µg/ml of Ly-6G (clone 1A8) Alexa Fluor® 594 (red), 2.5 µg/ml of CD3 (clone 145-2C11) Alexa Fluor® 647 (blue), and 2.5 µg/ml of B220 (clone RA3-6B2) Alexa Fluor® 488 (green) overnight at 4°C. The image was captured by 10X objective.
Compare all formats See Alexa Fluor® 594 spectral data
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127636 100 µg $293
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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

Product 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® 594 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

IHC-F - Quality tested
SB - Community verified
Recommended Usage

Each lot of this antibody is quality control tested by immunohistochemical staining on frozen tissue sections. For immunohistochemistry, a concentration range of 2.0 - 5.0 μg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 594 has an excitation maximum of 590 nm, and a maximum emission of 617 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

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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).

Additional Product Notes

This product has been verified for IHC-F (Immunohistochemistry - frozen tissue sections) and IHC-P (Immunohistochemistry - formalin-fixed paraffin-embedded tissues) on the NanoString GeoMx® Digital Spatial Profiler. The GeoMx® enables researchers to perform spatial analysis of protein and RNA targets in FFPE and fresh frozen human and mouse samples. For more information about our spatial biology products and the GeoMx® platform, please visit our spatial biology page.

View more applications data for this product in our Scientific Poster Library.

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. Choudhury SR, et al. 2020. Cell. 178(5):1205-1221.e17.. PubMed
  2. Qi JL, et al. 2020. MedComm (Beijing). 1:188. PubMed
  3. Ueki H, et al. 2020. Nat Protoc. 15:1041. PubMed
  4. Ouyang J, et al. 2020. Biomed Res Int. 4307385:2020. PubMed
  5. Pitsch J, et al. 2021. Ann Neurol. 89:666. PubMed
  6. Moniaga C, et al. 2015. Sci Rep. 5: 15319. PubMed
  7. Simões FC, et al. 2020. Nat Commun. 0.875. PubMed
  8. Li MY, et al. 2021. Developmental Cell. 56:2547. PubMed
  9. Chmielewski M and Abken H 2017. Cell Rep.. 10.1016/j.celrep.2017.11.063. PubMed
  10. Lee EKS, et al. 2018. Cell Host Microbe. 23:121. PubMed
  11. Wang F, et al. 2021. Oxid Med Cell Longev. 2021:5833857. PubMed
RRID
AB_2563207 (BioLegend Cat. No. 127636)

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.

Other Formats

View All Ly-6G Reagents Request Custom Conjugation
Description Clone Applications
Alexa Fluor® 594 anti-mouse Ly-6G 1A8 IHC-F,SB
Purified anti-mouse Ly-6G 1A8 FC,IHC-F,IHC-P,SB
Biotin anti-mouse Ly-6G 1A8 FC,IHC
FITC anti-mouse Ly-6G 1A8 FC
PE anti-mouse Ly-6G 1A8 FC
Alexa Fluor® 647 anti-mouse Ly-6G 1A8 FC,IHC-F,SB
Pacific Blue™ anti-mouse Ly-6G 1A8 FC
APC anti-mouse Ly-6G 1A8 FC
PerCP/Cyanine5.5 anti-mouse Ly-6G 1A8 FC
PE/Cyanine7 anti-mouse Ly-6G 1A8 FC
Alexa Fluor® 700 anti-mouse Ly-6G 1A8 FC,IHC
APC/Cyanine7 anti-mouse Ly-6G 1A8 FC
Alexa Fluor® 488 anti-mouse Ly-6G 1A8 FC,IHC-F,SB
Brilliant Violet 421™ anti-mouse Ly-6G 1A8 FC,IHC-F
Brilliant Violet 570™ anti-mouse Ly-6G 1A8 FC
Ultra-LEAF™ Purified anti-mouse Ly-6G 1A8 FC,Depletion,IHC
Brilliant Violet 510™ anti-mouse Ly-6G 1A8 FC
Purified anti-mouse Ly-6G (Maxpar® Ready) 1A8 FC,CyTOF®,WB
Brilliant Violet 650™ anti-mouse Ly-6G 1A8 FC
Brilliant Violet 711™ anti-mouse Ly-6G 1A8 FC
Brilliant Violet 605™ anti-mouse Ly-6G 1A8 FC
Brilliant Violet 785™ anti-mouse Ly-6G 1A8 FC
PE/Dazzle™ 594 anti-mouse Ly-6G 1A8 FC
APC/Fire™ 750 anti-mouse Ly-6G 1A8 FC
PerCP anti-mouse Ly-6G 1A8 FC
TotalSeq™-A0015 anti-mouse Ly-6G 1A8 PG
TotalSeq™-C0015 anti-mouse Ly-6G 1A8 PG
TotalSeq™-B0015 anti-mouse Ly-6G 1A8 PG
Spark Blue™ 550 anti-mouse Ly-6G 1A8 FC
Spark NIR™ 685 anti-mouse Ly-6G 1A8 FC
Spark YG™ 593 anti-mouse Ly-6G 1A8 FC
APC/Fire™ 810 anti-mouse Ly-6G Antibody 1A8 FC
PE/Cyanine5 anti-mouse Ly-6G 1A8 FC
PE/Fire™ 810 anti-mouse Ly-6G Antibody 1A8 FC
Spark UV™ 387 anti-mouse Ly-6G 1A8 FC
PE/Fire™ 640 anti-mouse Ly-6G 1A8 FC
Spark YG™ 570 anti-mouse Ly-6G 1A8 IHC-F,FC
Go To Top Version: 5    Revision Date: 01/24/2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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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.
If you need assistance with selecting the best format contact our expert technical support team.

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