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Purified 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
1-1A8_Pure_061708.jpg
C57BL/6 bone marrow cells stained with 1A8 purified, followed by anti-rat IgG PE (myeloid cells were gated for analysis)
  • 1-1A8_Pure_061708.jpg
    C57BL/6 bone marrow cells stained with 1A8 purified, followed by anti-rat IgG PE (myeloid cells were gated for analysis)
  • 2-1A8_Purified_Ly-6G_Antibody_IHC_102518
    Fresh, frozen mouse spleen was stained with purified Ly6G clone 1A8 conjugated and detected with a Cy5 CODEX™ oligonucleotide duplex (red). Samples were counterstained with TCR FITC (green). Data generated at Akoya Biosciences, Inc. using the CODEX™ technology.
  • 3-1A8_Purified_Ly-6G_Antibody_IHC_022013
    Mouse uterine tissue fixed in 10% formalin, paraffin embedded, and sliced to 4 µm. After deparaffination and antigen retrieval, sample was stained using an automatic slide stainer. The anti-mouse Ly6G primary antibody was applied at 1:500 dilution in blocking buffer for 1 hr at RT and DAB was used for visualization.
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127601 50 µg 52€
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127602 500 µg 111€
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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.
Concentration
0.5 mg/ml
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

FC - Quality tested
IHC-F - Verified
IHC - Reported in the literature, not verified in house
IHC-P, 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.

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-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.

Application References
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RRID
AB_1089179 (BioLegend Cat. No. 127601)
AB_1089180 (BioLegend Cat. No. 127602)

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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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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