TLRs belong to a family of proteins that specifically recognizes and senses microbial products. They are highly conserved throughout evolution and act as innate immune recognition receptors against many pathogens. TLR4 is a functional receptor for gram-negative bacterial lipopolysaccharides (LPS). TLR4 associates with MD-2 which is absolutely required for LPS-induced activation of TLR4. MD-2 exists as a cell surface protein in association with TLR4. It also exists as secreted forms consisting of MD-2 monomers and multimers (sMD-2). Circulating sMD-2 is mainly present as a doublet of ~20 and 25kD, representing differentially glycosylated forms. Unlike TLR4, sMD-2 binds directly LPS without the need of soluble CD14 (sCD14). However, LPS-MD-2 interactions are increased when LPS is pretreated with CD14. Only monomeric sMD-2 is biologically active and able to associate with TLR4 and LPS. sMD-2 circulates in plasma of healthy individuals as a non-active, polymeric protein. In septic plasma, the total amount of sMD-2 was strongly elevated and contained both sMD-2 polymers and monomers. Soluble MD-2 is proposed to be an important mediator of organ inflammation during sepsis. During experimental human endotoxemia, the monomeric and total sMD-2 content in plasma increased with the kinetics of an acute phase protein. This parallels enhanced TLR4 costimulatory activity. In vitro studies revealed that sMD-2 release appears to be restricted to endothelial and dendritic cells.
Applications:
Suitable for use in Flow Cytometry and Inhibition biological activity of the TLR4/MD-2 complex. Other applications not tested.
Recommended Dilution:
Flow Cytometry: 1:50
Optimal dilutions to be determined by the researcher.
Storage and Stability:
May be stored at 4 degrees C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20 degrees C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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