Alexa Fluor® 647 anti-mouse Ly-6G/Ly-6C (Gr-1) Antibody

Pricing & Availability
Clone
RB6-8C5 (See other available formats)
Regulatory Status
RUO
Other Names
Gr-1
Isotype
Rat IgG2b, κ
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Product Citations
publications
RB6-8C5_ALX647_020708
C57BL/6 mouse bone marrow (gated on myeloid cell population) stained with Ly-6G/Ly-6C (clone RB6-8C5) Alexa Fluor® 647 (filled histogram) or rat IgG2b, κ Alexa Fluor® 647 isotype control (open histogram).
  • RB6-8C5_ALX647_020708
    C57BL/6 mouse bone marrow (gated on myeloid cell population) stained with Ly-6G/Ly-6C (clone RB6-8C5) Alexa Fluor® 647 (filled histogram) or rat IgG2b, κ Alexa Fluor® 647 isotype control (open histogram).
Compare all formats See Alexa Fluor® 647 spectral data
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108420 25 µg £70
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108418 100 µg £157
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Description

Gr-1 is a 21-25 kD protein also known as Ly-6G/Ly-6C. This myeloid differentiation antigen is a glycosylphosphatidylinositol (GPI)-linked protein expressed on granulocytes and macrophages. In bone marrow, the expression levels of Gr-1 directly correlate with granulocyte differentiation and maturation; Gr-1 is also transiently expressed on bone marrow cells in the monocyte lineage. Immature Myeloid Gr-1+ cells play a role in the development of antitumor immunity.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Raised against granulocytes of mouse origin
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® 647 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
SB - Community verified

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 ≤ 0.25 µg per 106 cells in 100 µl volume. It is recommended that the reagent be titrated for optimal performance for each application.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633nm / 635nm.


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

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Excitation Laser
Red Laser (633 nm)
Application Notes

Clone RB6-8C5 binds with high affinity to mouse Ly-6G molecules and to a lower extent to Ly-6C19. Clone RB6-8C5 impairs the binding of anti-mouse Ly-6G clone 1A819. However, clone RB6-8C5 is able to stain in the presence of anti-mouse Ly-6C clone HK1.420.

The RB6-8C5 antibody has been used to identify peripheral blood neutrophils and deplete granulocytes in vivo. Additional reported applications (for relevant formats of this clone) include: in vitro complement-mediated cytotoxicity2, in vivo depletion3-5,9, immunoprecipitation1, immunohistochemical staining6 (including paraffin-embedded sections9,16,33-35, acetone-fixed frozen sections11 and zinc-fixed sections15), and Western blotting7. RB6-8C5 is not suitable for depletion of hepatic myeloid derived suppressor cells (MDSCs)20.

Special Note: For in vivo studies or highly sensitive assays, we recommend Ultra-LEAF™ purified antibody (Cat. No. 108436).

Additional Product Notes

This product has been verified for IHC-F (Immunohistochemistry - frozen tissue sections) 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.

Application References

(PubMed link indicates BioLegend citation)
  1. Fleming TJ, et al. 1993. J. Immunol. 151:2399. (IP)
  2. Brummer E, et al. 1984. J. Leukocyte Biol. 36:505. (CMCD)
  3. Stoppacciaro A, et al. 1993. J. Exp. Med. 178:151. (Deplete)
  4. Tumpey TM, et al. 1996. J. Virol. 70:898. (Deplete)
  5. Czuprynski CJ, et al. 1994. J. Immunol. 152:1836. (Deplete)
  6. Nitta H, et al. 1997. Cell Vision 4:73. (IHC)
  7. Jutila MA, et al. 1988. Eur. J. Immunol. 18:1819. (WB)
  8. Engwerda CR, et al. 2004. Am. J. Pathol. 165:2123.
  9. Brown CR, et al. 2004. Infect. Immun. 72:4956. (Deplete, IHC)
  10. Andoniou CE, et al. 2005. Nature Immunology 6:1011. (FC) PubMed
  11. Li M, et al. 2006. P. Natl. Acad. Sci USA 103:11736. (IHC)
  12. Dzhagalov I, et al. 2007. Blood 109:1620. (FC) PubMed
  13. Fazilleau N, et al. 2007. Nature Immunol. 8:753. (FC) PubMed
  14. Heuser M, et al. 2007. Blood 110:1639. (FC) PubMed
  15. Wang T, et al. 2007. Infect. Immun. 75:1144. (IHC)
  16. Bosio CM, et al. 2007. J. Immunol. 178:4538. (IHC)
  17. Boehme SA, et al. 2009. Int. Immunol. 21:81. (IHC)
  18. Piao Y, et al. 2012. Neuro Oncol. 14:1379. PubMed
  19. Ribechini E, et al. 2009. Eur. J. Immunol. 39:3538.
  20. Ma C, et al. 2012. J. Leukoc. Biol. 92:1199.
  21. Li J, et al. 2012. Arthritis Rheum. 64:1098. PubMed
  22. Fan Q, et al. 2014. Cancer Res. 74:471. PubMed
  23. Korrer MJ, et al. 2014. PLoS One. 9:91370. PubMed
  24. Morshed M, et al. 2014. J Immunol. 192:5314. PubMed
  25. Collins C, et al. 2014. PNAS. 111:9899. PubMed
  26. Madireddi S, et al. 2014. J Exp Med. 211:1433. PubMed
  27. Bianchi G, et al. 2014. Cell Death Dis. 5:1135. PubMed
  28. Guo H, et al. 2014. J Leukoc Biol. 96:419. PubMed
  29. Roderick JE, et al. 2014. PNAS. 111:14436. PubMed
  30. Distel E, et al. 2014. Circ Res. 115:759. PubMed
  31. Iwai H, et al. 2015. Tuberculosis. 95:246. PubMed
  32. Charmsaz S, et al. 2015. PLoS One. 10:130692. PubMed
  33. Whiteland J, et al. 1994 J Histochem Cytochem 43:3 (IHC-P)
  34. Brown C, et al. 2003 J Immunology 171:2 (IHC-P)
  35. Obregon-Henao A, et al. PLoS One 8:11 (IHC-P)
Product Citations
  1. Tan L, et al. 2022. Biochem Biophys Rep. 32:101351. PubMed
  2. Tarban N, et al. 2022. Cells. 11:. PubMed
  3. Wang J, et al. 2012. Blood. 120:1489. PubMed
  4. Ni J, et al. 2022. Mol Med. 28:65. PubMed
  5. Demars A, et al. 2021. PLoS Pathog. 17:e1009887. PubMed
  6. Shaw O, Harper J 2011. Biochem Biophys Res Commun. 416:266. PubMed
  7. Zhang H, et al. 2017. Leukemia. 10.1128/mBio.00226-17. PubMed
  8. Koliaraki V et al. 2019. Cell reports. 26(3):536-545 . PubMed
  9. Bittner–Eddy PD, et al. 2017. Front Immunol. 1.304166667. PubMed
  10. Chen G, et al. 2015. Sci Rep. 5: 14780. PubMed
  11. Meraz I, et al. 2014. PLoS One. 9:94703. PubMed
  12. Wheeler R, Gull E 2015. Nat Commun. 6: 8964. PubMed
  13. Bouchery T, et al. 2020. Cell Host & Microbe. 27(2):277-289. PubMed
  14. Biburger M, Nimmerjahn F 2012. Immunol Lett. 143:53. PubMed
  15. Tummers B, et al. 2020. Immunity. 52(6):994-1006.e8. PubMed
  16. Kim K, et al. 2013. J Vis Exp. 74: 50329. PubMed
  17. Budai Z, et al. 2021. Cells. 10:. PubMed
  18. Hahm E, et al. 2016. Nat Med. 23:100-106. PubMed
  19. Hendrikx S et al. 2019. Cell reports. 26(5):1227-1241 . PubMed
  20. Rossnagl S, et al. 2016. PLoS Biol. 14: 1002562. PubMed
  21. Al-Zaeed N, et al. 2021. Cell Death Dis. 12:611. PubMed
RRID
AB_493481 (BioLegend Cat. No. 108420)
AB_389331 (BioLegend Cat. No. 108418)

Antigen Details

Structure
21-25 kD
Distribution

Granulocytes, monocytes

Cell Type
Granulocytes, Monocytes, Neutrophils
Biology Area
Immunology, Innate Immunity
Antigen References

1. Fleming TJ, et al. 1993. J. Immunol. 151:2399.
2. Jutila MA, et al. 1988. Eur. J. Immunol. 18:1819.
3. Goni O, et al. 2002. Int. Immunol. 14:1125.

Gene ID
17067 View all products for this Gene ID 546644 View all products for this Gene ID
UniProt
View information about Ly-6G Ly-6C 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/Ly-6C Reagents Request Custom Conjugation
Description Clone Applications
APC anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Biotin anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC,IHC,IP,WB
FITC anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
PE anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
PE/Cyanine5 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Purified anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC,IHC,IP,WB
PE/Cyanine7 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Alexa Fluor® 488 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC,IHC
Alexa Fluor® 647 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC,SB
Alexa Fluor® 700 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Brilliant Violet 711™ anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
APC/Cyanine7 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Pacific Blue™ anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
PerCP/Cyanine5.5 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
PerCP anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Brilliant Violet 421™ anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Brilliant Violet 570™ anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Ultra-LEAF™ Purified anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC,IP,CMCD,Depletion,IHC,WB
Brilliant Violet 510™ anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Brilliant Violet 605™ anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Brilliant Violet 650™ anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Alexa Fluor® 594 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 IHC-F,FC
Purified anti-mouse Ly-6G/Ly-6C (Gr-1) (Maxpar® Ready) RB6-8C5 FC,CyTOF®
PE/Dazzle™ 594 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
APC/Fire™ 750 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
TotalSeq™-A0116 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 PG
TotalSeq™-C0116 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 PG
TotalSeq™-B0116 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 PG
Spark Blue™ 550 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
APC/Fire™ 810 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Spark Violet™ 423 anti-mouse Ly-6G/Ly-6C (GR-1) Antibody RB6-8C5 FC
Spark UV™ 387 anti-mouse Ly-6G/Ly-6C (GR-1) RB6-8C5 FC
Spark Violet™ 538 anti-mouse Ly-6G/Ly-6C (Gr-1) RB6-8C5 FC
Go To Top Version: 4    Revision Date: 01/24/2024

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

 

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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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