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 Alexa Fluor® 647 conjugated products with eFluor® 660 conjugated products.
Description
The UT41 monoclonal antibody was generated against BaF3 cells overexpressing mouse TLR4. So far, at least twelve members of the Toll family have been identified. This family of type I transmembrane protein 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 another molecule called MD-2, and together with CD14, this complex is responsible for LPS recognition and signaling. TLR4, which is approximately 100 kDa, is expressed by myeloid lineage cells.
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 UT41 antibody has been reported for use in flow cytometric analysis.
Applications Tested
This UT41 antibody has been tested by flow cytometric analysis of mouse 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 105 to 108 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.
References
Nomura, F., S. Akashi, et al. 2000. Cutting edge: endotoxin tolerance in mouse peritoneal macrophages correlates with down-regulation of surface toll-like receptor 4 expression. J Immunol 164(7): 3476-9.
Sato, S., F. Nomura, et al. 2000. Synergy and cross-tolerance between toll-like receptor (TLR) 2- and TLR4-mediated signaling pathways. J Immunol 165(12): 7096-101.