Ly-6G/Ly-6C Antibody (RB6-8C5) - Chimeric - Azide and BSA Free

Novus Biologicals | Catalog # NBP3-11978

Recombinant Monoclonal Antibody
Novus Biologicals
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Key Product Details

Species Reactivity

Mouse

Applications

Western Blot, Flow Cytometry, Immunocytochemistry/ Immunofluorescence

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Kappa Clone # RB6-8C5

Format

Azide and BSA Free
View all Ly-6G (Gr-1) Primary Antibodies »

Product Specifications

Immunogen

This antibody was raised in Lou-M rats using a popliteal lymph node immunisation and fusion protocol

Specificity

This antibody is specific for murine Ly6G, a a 21-25 kDa glycosylphosphatidylinositol (GPI)-linked protein expressed by the myeloid-derived cells in a tightly developmentally-regulated manner in the bone marrow. Monocytes express Ly6G transiently during bone marrow development, while Ly6G expression in granulocytes and peripheral neutrophils directly correlates with the cell's level of differentiation and maturation. This hallmark makes Ly6G a good marker for these particular cell populations. Ly6G has also been implicated in the development of antitumor responses.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG Kappa

Scientific Data Images for Ly-6G/Ly-6C Antibody (RB6-8C5) - Chimeric - Azide and BSA Free

Immunocytochemistry/ Immunofluorescence: Ly-6G/Ly-6C Antibody (RB6-8C5) - Chimeric - Azide and BSA Free [NBP3-11978]

Immunocytochemistry/ Immunofluorescence: Ly-6G/Ly-6C Antibody (RB6-8C5) - Chimeric - Azide and BSA Free [NBP3-11978]

Immunocytochemistry/Immunofluorescence: Ly-6G/Ly-6C Antibody (RB6-8C5) - Chimeric [NBP3-11978] - Immunofluorescence analysis of paraformaldehyde fixed mouse splenocytes immobilized on Shi-fix(TM) cover-slips and stained with the chimeric rabbit IgG version of RB6-8C5 (NBP3-11978) at 10 ug/ml followed by Alexa Fluor(R) 488 secondary antibody (2 ug/ml), showing membrane staining in subset of cells. The nuclear stain is DAPI (blue). Panels show from left-right, top-bottom NBP3-11978, DAPI, merged channels and an isotype control. The isotype control was stained with anti-Fluorescein antibody (NBP2-52638) followed by Alexa Fluor(R) 488 secondary antibody.
Flow Cytometry: Ly-6G/Ly-6C Antibody (RB6-8C5) - Chimeric - Azide and BSA Free [NBP3-11978]

Flow Cytometry: Ly-6G/Ly-6C Antibody (RB6-8C5) - Chimeric - Azide and BSA Free [NBP3-11978]

Flow Cytometry: Ly-6G/Ly-6C Antibody (RB6-8C5) - Chimeric [NBP3-11978] - Mouse splenocytes were stained with anti-Fluorescein IgG antibody (4-4-20; isotype control, black line) or the rabbit IgG version of RB6-8C5 (NBP3-11978, blue line) at a dilution of 1:100 for 1h at RT. After washing, bound antibody was detected using a goat anti-rabbit IgG AlexaFluor(R) 488 antibody at a dilution of 1:1000 and cells analyzed using a FACSCanto flow-cytometer.

Applications for Ly-6G/Ly-6C Antibody (RB6-8C5) - Chimeric - Azide and BSA Free

Application
Recommended Usage

Flow Cytometry

Optimal dilutions of this antibody should be experimentally determined.

Immunocytochemistry/ Immunofluorescence

Optimal dilutions of this antibody should be experimentally determined.

Western Blot

Optimal dilutions of this antibody should be experimentally determined.
Application Notes
This chimeric rabbit antibody was made using the variable domain sequences of the original Rat IgG2a format, for improved compatibility with existing reagents, assays and techniques.

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A purified

Formulation

PBS

Format

Azide and BSA Free

Preservative

0.02% Proclin 300

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C for up to 3 months. For longer storage, aliquot and store at -20C.

Background: Ly-6G (Gr-1)

The lymphocyte antigen 6 (Ly-6) complex was first described in mice and since its discovery over 20 genes related to the Ly-6 complex on chromosome 15 have been classified (1). The Ly-6 family is commonly used as a cell-surface differentiation marker on both normal and abnormal cells (2). Ly-6 complex, locus G (Ly-6G) and Ly-6 complex, locus C (Ly-6C) both belong to the murine Ly6/urokinase-type plasminogen activator receptor (uPAR) family of genes, which share a common structure (3). The Ly-6 family of proteins is characterized by a ~90 amino acid (aa) LU domain that consists of a three-finger structure formed by disulfide bonds (1,3). Additionally, many Ly6 proteins have a C-terminal GPI anchor, including Ly-6G/Ly-6C (1,3). Ly-6C protein has a molecular weight of 14 kDa and is expressed in some CD4+ and CD8+ T-cells, monocytes, neutrophils, and dendritic cells (1). Ly-6C has uniform expression in neutrophils but varies in monocytes where Ly-6Chi expression is specific for immature/inflammatory monocytes and Ly-6Clo expression identifies mature/residential monocytes (1,3). Ly-6G, on the other hand, is a protein with a molecular weight of 25 kDa with its expression uniquely restricted to neutrophils (1).

Expression of Ly-6G and Ly-6C, along with other lineage markers, help define specific cell types (2,4). For example, mouse myeloid derived suppressor cells (MDSCs), which play a role in inflammation and tumor development, are further subdivided into granulocytic (G-MDSCs) or monocytic (Mo-MDSCs) (4). Mo-MDSCs express Ly-6Chi along with CD11b, but are negative for Ly-6G, whereas G-MDSCs express Ly-6Clo/int,Ly-6Ghi, and CD11b (4).

The monoclonal antibody RB6-8C5 first identified granulocyte-differentiation antigen (Gr-1) which is primarily expressed on the surface of neutrophils and immature myeloid cells, and to a lesser extent on lymphocytes and macrophages (1). Gr-1 expression increases as neutrophils mature but remains is stably expressed in monocytes (1). The RB6-8C5 reacts with both Ly-6G and Ly-6C, whereas 1A8 antibody is specific for Ly-6G (1). Antibodies against Ly-6G, such as RB6-8C5 and 1A8, are commonly used to identify a role for neutrophils in mouse models of disease as they are efficient for quickly depleting neutrophils (1). RB6-8C5, however, also targets and depletes Ly-6Chi monocytes in addition to other Ly-6G-expressing cells (1).

References

1. Lee, P. Y., Wang, J. X., Parisini, E., Dascher, C. C., & Nigrovic, P. A. (2013). Ly6 family proteins in neutrophil biology. Journal of leukocyte biology. https://doi.org/10.1189/jlb.0113014

2. Bamezai A. (2004). Mouse Ly-6 proteins and their extended family: markers of cell differentiation and regulators of cell signaling. Archivum immunologiae et therapiae experimentalis.

3. Loughner, C. L., Bruford, E. A., McAndrews, M. S., Delp, E. E., Swamynathan, S., & Swamynathan, S. K. (2016). Organization, evolution and functions of the human and mouse Ly6/uPAR family genes. Human genomics. https://doi.org/10.1186/s40246-016-0074-2

4. Kong, Y. Y., Fuchsberger, M., Xiang, S. D., Apostolopoulos, V., & Plebanski, M. (2013). Myeloid derived suppressor cells and their role in diseases. Current medicinal chemistry. https://doi.org/10.2174/0929867311320110006

Long Name

A Myeloid Differentiation Antigen

Alternate Names

Gr-1

Gene Symbol

LY6G

Additional Ly-6G (Gr-1) Products

Product Documents for Ly-6G/Ly-6C Antibody (RB6-8C5) - Chimeric - Azide and BSA Free

Certificate of Analysis

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Product Specific Notices for Ly-6G/Ly-6C Antibody (RB6-8C5) - Chimeric - Azide and BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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