Staining of 3-day thioglycolate-induced BALB/c macrophages with 0.25 µg of Rat IgG2a kappa Isotype Control PE (Product # 12-4321) (open histogram) or 0.25 µg of Anti-Mouse TLR4/MD-2 Complex PE (filled histogram). Cells were pre-incubated with Anti-Mouse CD16/CD32 Purified (Product # 14-0161). Total viable cells were used for analysis.
|Tested species reactivity||Mouse|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rat / IgG2a, kappa|
|Storage buffer||PBS, pH 7.2, with 0.1% gelatin|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||0.5 µg/test|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
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: MTS510 has been reported for use in flow cytometric analysis.
Applications Tested: The MTS510 antibody has been tested by flow cytometric analysis of mouse thioglycolate-elicited peritoneal exudate cells. This can be used at less than or equal to 0.5 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser
MD-2 is a member of the Toll/interleukin-1 receptor (TIR) family, a group of proteins that include the Toll-like receptors (TLRs). TLRs are signaling molecules that recognize different pathogen-associated molecular patterns (PAMPs) and serve as an important link between the innate and adaptive immune responses. TLR4, the major signaling receptor for lipopolysaccharide (LPS), requires the binding of MD-2 to its extracellular region for maximal response to LPS. The specificity of this response is determined by the species of MD-2; e.g., human MD-2 transfected into mouse cells can cause mouse TLR4 to react to LPS analogs that are normally antagonistic to human but not mouse TLR4.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
||TLRs regulate the gatekeeping functions of the intestinal follicle-associated epithelium.||Chabot S,Wagner JS,Farrant S,Neutra MR||Journal of immunology (Baltimore, Md. : 1950) (176:4275)||2006|
|Not Applicable||Not Cited||TLRs regulate the gatekeeping functions of the intestinal follicle-associated epithelium.||Chabot S,Wagner JS,Farrant S,Neutra MR||Journal of immunology (Baltimore, Md. : 1950) (176:4275)||2006|