Product Information

Contents: Affinity Purified anti-mouse MD-1
Catalog Number: 14-9921
Sizes: 50 ug, 100 ug
Formulation: Phosphate buffer pH 7.2,
150 mM NaCl, 0.09% NaN₃
Storage Conditions: Store at 4°C.
Avoid repeated freeze/thaw cycles.
Clone: MD14
Isotype: Rat IgG2a, κ

Surface staining of mouse splenocytes with anti-mouse MD-1 (MD14) PE. Appropriate isotype controls were used (open histogram). Total viable cells were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-9921	Affinity Purified anti-mouse MD-1 (MD1; Toll-like Receptor/ TLR Family)	N/A	N/A	FC
13-9921	Biotin anti-mouse MD-1 (MD1; Toll-like Receptor/ TLR Family)	N/A	N/A	FC
12-9921	PE anti-mouse MD-1 (MD1; Toll-like Receptor/ TLR Family)	488	575	FC
Flow Cytometry Product Notes: Test Sizes: To accommodate multicolor flow cytometry, eBioscience is in the process of reducing test size volumes from 20 µl to 5 µl. Please check your antibody vial for the recommended test size. Fluorochrome Replacements: eBioscience is in the process of replacing all Pacific Blue® and APC-Alexa Fluor® 750 conjugated products with eFluor™ 450 and APC-eFluor™ 780 conjugated products, respectively.

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Description

The MD14 monoclonal antibody reacts with mouse MD-1, a 28 kDa molecule, which physically associates with the extracellular portion of RP105 (CD180). MD-1 is expressed by mature B cells, monocytes/macrophages and dendritic cells. The coexpression of MD-1 is indispensable for cell surface expression of RP105 and LPS recognition and signaling. Several monoclonal antibodies to mouse MD-1, including MD14 and MD113, have been reported to be antagonistic for LPS-induced B cell proliferation and CD86 upregulation, while the anti-mouse CD180 (clone RP/14, Cat. No. 16-1801) is reported to exert potent mitogenic effect. The mAb MD113 (Cat. No. 16-9931) is reported to suppress LPS-induced B cell responses to a higher degree than MD14.

Applications Reported

For research use only, not for diagnostic or therapeutic use. MD14 has been reported for use in flow cytometric analysis. MD-14 is not useful for immunoprecipitation and immunoblotting (WB). Applicability of MD14 for use in immunohistochemistry has not been evaluated.

Applications Tested

The MD14 antibody has been tested by flow cytometric analysis of mouse spleen cell suspensions and can be used at less than or equal to 1 µg per million cells. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

References

Nagai, Y., R. Shimazu, et al. 2002. Requirement for MD-1 in cell surface expression of RP105/CD180 and B-cell responsiveness to lipopolysaccharide. Blood 99(5):1699-705.
Miyake, K., R. Shimazu, et al. 1998. Mouse MD-1, a molecule that is physically associated with RP105 and positively regulates its expression. J Immunol 161(3): 1348-53.
Miyake, K., H. Ogata, et al. 2000. Innate recognition of lipopolysaccharide by Toll-like receptor 4/MD-2 and RP105/MD-1. J Endotoxin Res 6(5):389-91.
Miura, Y., R. Shimazu, et al. 1998. RP105 is associated with MD-1 and transmits an activation signal in human B cells. Blood 92(8):2815-22.
Gorczynski, R. M., Z. Chen, et al. 2000. Regulation of gene expression of murine MD-1 regulates subsequent T cell activation and cytokine production. J Immunol 165(4):1925-32.
Ogata, H., I. Su, et al. 2000. The toll-like receptor protein RP105 regulates lipopolysaccharide signaling in B cells. J Exp Med 192(1):23-9.

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