Ly-6G/Ly-6C Antibody (RB6-8C5) - Azide and BSA Free Summary
Immunogen |
Normal mouse bone marrow. |
Isotype |
IgG2b Kappa |
Clonality |
Monoclonal |
Host |
Rat |
Gene |
Ly6g |
Purity |
Protein A or G purified |
Innovator's Reward |
Test in a species/application not listed above to receive a full credit towards a future purchase. |
Applications/Dilutions
Dilutions |
- CyTOF-ready
- Flow Cytometry 1:10-1:1000
- Immunocytochemistry/ Immunofluorescence 8-25 ug/ml
- Immunohistochemistry 1:10-1:500
- Immunohistochemistry-Frozen 1:10-1:500
- Immunohistochemistry-Paraffin
- Immunoprecipitation 1:10-1:500
- In vivo assay reported in scientific literature (PMID 16272176)
- Western Blot 1:100-1:2000
|
Application Notes |
The RB6-8C5 antibody has been tested by flow cytometric analysis of mouse bone marrow cells and splenocyte suspensions. This can be used at less than or equal to 0.5 ug per test. A test is defined as the amount (ug) of antibody that will stain a cell sample in a final volume of 100 ul. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. The RB6-8C5 antibody has also been reported for use immunoprecipitation, immunoblotting (WB) and immunohistochemical staining. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. This antibody is CyTOF ready. |
Packaging, Storage & Formulations
Storage |
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. |
Buffer |
PBS |
Preservative |
No Preservative |
Concentration |
1 mg/ml |
Purity |
Protein A or G purified |
Alternate Names for Ly-6G/Ly-6C Antibody (RB6-8C5) - Azide and BSA Free
Background
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
Limitations
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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