Product Information

Contents: Affinity Purified anti-mouse Toll-like receptor 4 (TLR4)/MD-2
Catalog Number: 14-9924
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: MTS510
Isotype: Rat IgG2a, κ

Surface staining of thioglycolate induced PEC with anti-mouse Toll-like receptor 4/MD-2 (MTS510) PE. Appropriate isotype controls were used (open histogram). Cells in the large scatter population were used for analysis.

Available Formats of This Product
Cat. No.	Format	Excite (nm)	Emit (nm)	Reported Applications
14-9924	Affinity Purified anti-mouse Toll-like receptor 4/ MD-2 (TLR4/ MD-2, TLR-4/ MD2)	N/A	N/A	FA FC IH/F IP
16-9924	Functional Grade* Purified anti-mouse Toll-like receptor 4/ MD-2 (TLR4/ MD-2, TLR-4/ MD2)	N/A	N/A	FA FC
13-9924	Biotin anti-mouse Toll-like receptor 4/ MD-2 (TLR4/ MD-2, TLR-4/ MD2)	N/A	N/A	FC
12-9924	PE anti-mouse Toll-like receptor 4/ MD-2 (TLR4/ MD-2, TLR-4/ MD2)	488	575	FC
17-9924	Allophycocyanin (APC) anti-mouse Toll-like receptor 4/ MD-2 (TLR4/ MD-2, TLR-4/ MD2)	633	660	FC
25-9924	Phycoerythrin-Cy7 (PE-Cy7) anti-mouse Toll-like receptor 4/ MD-2 (TLR4/ MD-2, TLR-4/ MD2)	488	760	FC
*Functional Grade™ (FG™) Purified: Azide-free, sterile-filtered, and endotoxin < 0.001 ng/µg (unless otherwise noted). *Functional Grade™ (FG™) Biotin: Azide-free, sterile-filtered, and endotoxin < 0.05 ng/µg. Purified: Contains azide, not sterile-filtered, and not endotoxin tested.
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 MTS510 monoclonal antibody reacts with the mouse Toll-like receptor 4 (TLR4)/MD-2 complex. At least ten members of the Toll family have been identified. This family of type I transmembrane proteins is characterized by an extracellular domain with leucine-rich repeats and a cytoplasmic domain with homology to the type I IL-1 receptor. Two of these receptors, TLR2 and TLR4, are pattern recognition receptors and signaling molecules in response to bacterial lipoproteins and have been implicated in innate immunity and inflammation. TLR4 physically associates with MD-2, and together with CD14, this complex is responsible for LPS recognition and signaling. In the mouse, TLR4 is expressed by thioglycolate-elicited peritoneal macrophages. Incubation of peritoneal macrophages with LPS results in down regulation of surface TLR4/MD-2. The TLR4 gene is defective in C3H/HeJ and C57BL/10ScCr mice, both of which have been well characterized as hyporesponders to LPS.

The MTS510 monoclonal antibody co-immunoprecipitates MD-2 (~30 kDa) and TLR4 (~100 kDa), and preferentially reacts with TLR4 that is associated with MD-2. In comparison, binding of the UT41 monoclonal antibody occurs with and without formation of the TLR4/MD-2 complex . Please contact eBioscience Technical Support for further information.

Applications Reported

For research use only, not for diagnostic or therapeutic use. This MTS510 antibody has been reported for use in flow cytometric analysis, immunoprecipitation, immunohistology staining of frozen tissue sections and inhibition of LPS-induced cytokine production. (Please use Functional Grade purified MTS510, cat. 16-9924, in functional assays.)

Applications Tested

The MTS510 antibody has been tested by flow cytometric analysis of mouse peritoneal exudate cells. This can be used at less than or equal to 1 μg per million cells in a 100 μl total staining volume. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

References

Chabot S, Wagner JS, Farrant S, Neutra MR. TLRs regulate the gatekeeping functions of the intestinal follicle-associated epithelium. J Immunol. 2006 Apr 1;176(7):4275-83. (MTS510, IHC frozen, PubMed)

Mortaz E, Redegeld FA, Nijkamp FP, Wong HR, Engels F. Acetylsalicylic acid-induced release of HSP70 from mast cells results in cell activation through TLR pathway. Exp Hematol. 2006 Jan;34(1):8-18. (MTS510, FA, PubMed)

Qi HY, Shelhamer JH. Toll-like receptor 4 signaling regulates cytosolic phospholipase A2 activation and lipid generation in lipopolysaccharide-stimulated macrophages. J Biol Chem. 2005 Nov 25;280(47):38969-75. (MTS510, FA, PubMed)

Ortega-Cava CF, Ishihara S, Rumi MA, Kawashima K, Ishimura N, Kazumori H, Udagawa J, Kadowaki Y, Kinoshita Y. Strategic compartmentalization of Toll-like receptor 4 in the mouse gut. J Immunol. 2003 Apr 15;170(8):3977-85. (MTS510, IHC frozen, PubMed)

Sato S, Nomura F, Kawai T, Takeuchi O, Mühlradt PF, Takeda K, Akira S. Synergy and cross-tolerance between toll-like receptor (TLR) 2- and TLR4-mediated signaling pathways. J Immunol. 2000 Dec 15;165(12):7096-101.

Akashi S, Shimazu R, Ogata H, Nagai Y, Takeda K, Kimoto M, Miyake K. Cutting edge: cell surface expression and lipopolysaccharide signaling via the toll-like receptor 4-MD-2 complex on mouse peritoneal macrophages. J Immunol. 2000 Apr 1;164(7):3471-5.

Nomura F, Akashi S, Sakao Y, Sato S, Kawai T, Matsumoto M, Nakanishi K, Kimoto M, Miyake K, Takeda K, Akira S. Cutting edge: endotoxin tolerance in mouse peritoneal macrophages correlates with down-regulation of surface toll-like receptor 4 expression. J Immunol. 2000 Apr 1;164(7):3476-9.

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