sMD-2 and sMD-2/TLR4, Human, mAb 4H1 - HM2243

Name

sMD-2 and sMD-2/TLR4, Human, mAb 4H1

Catalog nr

HM2243 (lot number and expiry date are indicated on the label)

Short description

The monoclonal antibody 4H1 reacts with soluble MD-2 (sMD-2) and the sMD-2/Toll-like receptor 4 (TLR4, CD284) complex. 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 lipo...

Size

100 µg

Application

FS , IA , IF , W

Technical datasheet

HM2243.pdf

Description

MD-2 exists as a cell surface protein in association with TLR4. It also exists as secreted forms consisting of MD-2 monomer and multimers. Circulating sMD-2 is mainly present as a doublet of ~20 and 25 kD, 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.

The monoclonal antibody 4H1 reacts with both the monomeric and the polymeric form of sMD-2. In addition, the monoclonal antibody 4H1 is able to inhibit LPS binding to MD-2.It does not cross-react with mouse MD-2.

Cross Reactivity

Cross reactant	Reactivity
Human	Yes

Immunogen

Baculovirally expressed His-tagged human MD-2

Formulation

1 ml (100 µg/ml) 0.2 µm filtered antibody solution in PBS, containing 0.1% bovine serum albumin.

Species

Mouse IgG1

Application

The monoclonal antibody 4H1 can be used for immuno assay, Western blotting and immunofluorescence. Furthermore, the antibody can be used to inhibit LPS binding to MD-2 in vitro.

Use

For immuno assay, Western blotting and immunofluorescence, dilutions to be used depend on detection system applied. It is recommended that users test the reagent and determine their own optimal dilutions. The typical starting working dilution is 1:50. For functional studies, dilutions have to be made according to the amounts of MD-2 to be inactivated.

Aliases

Lymphocyte antigen 96, ESOP-1, LY96

Storage and stability

Product should be stored at 4°C. Under recommended storage conditions, product is stable for one year.

References

Viriyakosol, S et al; Characterization of monoclonal antibodies to human soluble MD-2 protein. Hybridoma 2006, 25: 349

Precautions

For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use or derivation of this product.

Also available

HM2243F	sMD-2 and sMD-2/TLR4, Human, mAb 4H1, FITC
HM2244	sMD-2, human, mAb 10D5
HM2245	MD-2, Human, mAb 18H10
HM2246	TLR4/MD-2, Human, mAb 7E3

References

Viriyakosol, S et al; Characterization of monoclonal antibodies to human soluble MD-2 protein. Hybridoma 2006, 25: 349

Scientific info

Useful new antibodies for studies of MD-2 expression and function in health and disease

Hycult Biotech introduces four monoclonal antibodies to study both soluble and membrane bound human MD-2. MD-2 is a 20-25 kD glycoprotein that associates with the extracellular portion of Toll-like receptor 4 (TLR4, CD284). TLR4 association with MD-2 is absolutely required for LPS-induced activation of TLR4.

MD-2 also exists in secreted forms consisting of MD-2 monomers and multimers. Only monomeric soluble MD-2 (sMD-2) is biologically active and able to associate with TLR4 and LPS. sMD-2 circulates in plasma of healthy individuals as non-active, polymeric protein. In septic plasma, the total amount of sMD-2 was strongly elevated and contained both sMD-2 monomers and polymers.

sMD-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.

The four antibodies Hycult Biotech introduces here are reactive with MD-2 in its various forms. Three of them are useful for studies on biological activity of MD-2. All four monoclonal antibodies are available as biotinylated and FITC-labeled conjugates.

Cat. No.	Product	MD-2	sMD-2	MD-2/TLR4 complex	Inhibition of biological activity	Other applications
HM2243	sMD-2 and sMD-2/TLR4, human, mAb 4H1	+	+	+	+	IA IF W
HM2244	sMD-2, human, mAb 10D5	-	+	-	-	IA IF W
HM2245	MD-2, human, mAb 18H10	+	-	+	+	FC
HM2246	TLR4/MD-2, human, mAb 7E3	-	-	+	+	FC

References:
1. Viriyakosol, S et al; Characterization of monoclonal antibodies to human soluble MD-2 protein. Hybridoma 2006, 25: 349
2. Pugin, J et al; Soluble MD-2 activity in plasma from patients with severe sepsis and septic shock. Blood 2004, 104: 4071
3. Daubeuf, B et al; TLR4/MD-2 monoclonal antibody therapy affords protection in experimental models of septic shock. J Immunol 2007, 179: 6107
4. 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
5. Tissieres, P et al; Soluble MD-2 is an acute-phase protein and an opsonin for Gram-negative bacteria. Blood 2008, 111: 2122

MSDS

MSDS antibodies without azide.pdf

Protocols

Protocol Flow cytometry.pdf

Protocols

Protocol Immunoprecipitation.pdf

Protocols

Protocol Sandwich ELISA.pdf

Related products		Cat #
sMD-2, human, mAb 10D5		HM2244
TLR4/MD-2, Human, mAb 7E3		HM2246
MD-2, Human, mAb 18H10		HM2245