Technical Data

M2767-20P
Clone Type
Monoclonal
Host
Mouse
Source
Human
Conjugate
FITC
Isotype
IgG2b
Clone Number
13A58
Grade
Affinity Purified
Applications
FC
Crossreactivity
Hu
Shipping Temp
Blue Ice
Storage Temp
-20°C
Mouse Anti-MD-2 (Lymphocyte Antigen 96, Ly-96, ESOP-1, Protein MD-2, LY96, ESOP1) (FITC)

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. 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°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20°C. Aliquots are stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. FITC conjugates are sensitive to light.
Immunogen
Chimeric TLR4/MD-2 fusion protein expressed in CHO cells.
Form
Supplied as a liquid in PBS, 1% BSA, 0.02% sodium azide. Labeled with fluorescein isothiocyanate (FITC).
Purity
Purified by Protein G affinity chromatography.
Specificity
Recognizes human MD-2. Does not react with sMD-2. It inhibits bacterial binding to MD-2.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

References
1. Pugin, J et al; Soluble MD-2 activity in plasma from patients with severe sepsis and septic shock. Blood 2004, 104: 4071. 2. Daubeuf, B et al; TLR4/MD-2 monoclonal antibody therapy affords protection in experimental models of septic shock. J Immunol 2007, 179: 6107. 3. Elson, G et al; Contribution of Toll-like receptors to the innate immune response to Gram-negative and Gram-positive bacteria. Blood 2007, 109: 1574. 4. Tissieres, P et al; Soluble MD-2 is an acute phase protein and an opsonin for Gram-negative bacteria. Blood 2008, 111: 2122.
USBio References
No references available
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