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: This MTS510 antibody has been reported for use in flow cytometric analysis.
Applications Tested: This MTS510 antibody has been tested by flow cytometric analysis of mouse peritoneal-exudate cells. This can be used at less than or equal to 2 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.
Light sensitivity: This tandem dye is sensitive photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL cell sample + 100 µL IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
Filtration: 0.2 µm post-manufacturing filtered.
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.
Protein Aliases: CD284; ESOP-1; lipopolysaccharide response; Ly-96; Lymphocyte antigen 96; myeloid differentiation factor-2; myeloid differentiation protein-2; Protein MD-2; Toll-like receptor 4
Gene Aliases: ESOP-1; Esop1; Lps; Ly87; Ly96; MD-2; MD2; Ran/M1; Rasl2-8; Tlr4