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

Catalog # NBP2-00441 | Novus Biologicals a Bio-Techne Brand
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NBP2-00441-100ug
NBP2-00441SS

Key Product Details

Species Reactivity

Human, Mouse

Applications

CyTOF-ready, Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Immunohistochemistry-Frozen, Immunohistochemistry-Paraffin, Immunoprecipitation, In vivo assay, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2b Kappa Clone # RB6-8C5

Format

BSA Free

Concentration

1.0 mg/ml

Product Summary for Ly-6G/Ly-6C Antibody (RB6-8C5) - BSA Free

Immunogen

Normal mouse bone marrow.

Reactivity Notes

Use in Mouse reported in scientific literature (PMID:34449927).

Specificity

Studies indicate that in the bone marrow and lysed whole blood, the antibody clone RB6-8C5 also reacts with an additional subpopulation of Ly-6C high cells.

Clonality

Monoclonal

Host

Rat

Isotype

IgG2b Kappa

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

Western Blot: Ly-6G/Ly-6C Antibody (RB6-8C5) [NBP2-00441] - Mice were treated with oil in combination with injection of lenti-NC (NC, n = 10), CCl4 in combination with injection of lenti-NC (NC + CCl4, n = 10), oil in combination with injection of lenti-lnc-Lfar1-shRNA (lnc-Lfar1-shRNA, n = 10), and CCl4 in combination with injection of lenti-lnc-Lfar1-shRNA (lnc-Lfar1-shRNA + CCl4, n = 10). b The protein level of F4/80, CD11b and LY6C was determined by western blot. GAPDH was used as an internal control. Image collected and cropped by CiteAb from the following publication (http://www.nature.com/articles/s41419-020-2323-5), licensed under a CC-BY license.
Immunohistochemistry: Ly-6G/Ly-6C Antibody (RB6-8C5) [NBP2-00441] - Mice were treated with oil in combination with injection of lenti-NC (NC, n = 10), CCl4 in combination with injection of lenti-NC (NC + CCl4, n = 10), oil in combination with injection of lenti-lnc-Lfar1-shRNA (lnc-Lfar1-shRNA, n = 10), and CCl4 in combination with injection of lenti-lnc-Lfar1-shRNA (lnc-Lfar1-shRNA + CCl4, n = 10). a Immunohistochemistry analysis was performed to detect the expression of F4/80 and LY6C; scale bar = 400 um for 10 x and 100 um for 40x. Image collected and cropped by CiteAb from the following publication (http://www.nature.com/articles/s41419-020-2323-5), licensed under a CC-BY license.
Flow Cytometry: Ly-6G/Ly-6C Antibody (RB6-8C5) [NBP2-00441] - 1 ug/1^6 BALB/c bone marrow cells were stained with either rat IgG2b: FITC (as an isotype control) or rat anti-mouse Ly-6G: FITC. Large cells were then gated and analyzed on a flow cytometer.

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

Application
Recommended Usage

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

1:10-1:500

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.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 4.5 using NBP2-00441 in the following applications:

Published Applications

Read 24 publications using NBP2-00441 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified

Formulation

PBS

Format

BSA Free

Preservative

0.02% Sodium Azide

Concentration

1.0 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 short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

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

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

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

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

Showing  1 - 2 of 2 FAQs Showing All
  • Q: On the Datasheet of Ly-6G Antibody (RB6-8C5), Western Blot is listed as one of the applications. Do you have any suggestions on the concentration to use for this Ab in WB?

    A: We recommend dilutions between 1:1000 and 1:2000 as a starting point in Western blot.

  • Q: Can you please provide me with any publication reference the uses clone RB6-8C5 in Western?

    A: Unfortunately we do not have any references for use with this antibody in Western blot at the time.

Showing  1 - 2 of 2 FAQs Showing All
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