High quality reagents for innate immunity research, vaccine development and drug discovery

		Recombinant human LPS binding protein (LBP)
		Background Lipopolysaccharide Binding Protein (LBP) is a soluble protein present in plasma that is produced mainly by hepatocytes in the liver. The primary function of LBP is to transfer monomers of lipopolysaccharide (LPS, endotoxin) from larger aggregates, such as Gram-negative bacterial cell membranes or outer membrane vesicles, to the CD14 receptor on monocytes and macrophages [1]. It is an acute phase protein, reaching relatively high concentrations in plasma (e.g. up to 100 μg/ml in man) during periods of systemic inflammatory insult. It is thought this may contribute to a negative feedback response to excessive inflammation induced by endotoxaemia, since high concentrations of LBP can limit inflammatory cytokine production in response to LPS [1,2]. It associates loosely in the circulation with various lipoprotein particles (such as high density lipoprotein), so modifying their capacity to sequester LPS from detection by pattern-recognition receptors [3]. Caithness Biotech recombinant LBP comprises amino acids Ala 26 to Val 481 of the native human sequence. Potential applications of LBP protein include use in bioassays to modify cellular sensitivity to LPS, studies of the capacity of LBP to transfer LPS and other lipids to various acceptors, and assays requiring monomeric display of LPS polysaccharides in a protein-bound form (e.g. as a capture reagent for ELISA).
		Description
		Product	Recombinant human LPS binding protein (LBP)
		Size / volume	20 μg
		Expression system	HEK-293 cells
		Amino acids	Ala 26 to Val 481, accession number P18428
		Tags	C-terminal 6x His tag
		Sequence graphic
		Intended use	For laboratory research only, not for clinical or diagnostic use.
		Specifications
		Format	Lyophilised from sterile PBS (pH 7.4) with trehalose as protectant and without additional carrier protein.
		Purity	>95% by SDS PAGE
		Molecular weight	Migrates at ~ 63 kDa (glycosylation present)
		LPS content	< 0.1 ng / μg (by HEK-293-TLR4 bioassay, relative to E. coli LPS standard)
		BLP content	< 0.1 ng / μg (by HEK-293-TLR2 bioassay, relative to Pam3CSK4 standard)
		Amino acid sequence	See Technical data sheet.
		Applications	ELISA / bioassay / SDS PAGE / binding studies / immunoassays
		Pricing
		Catalogue number	LBP-1
		Price per unit	£100 per vial of 20 μg
		Figure 1: SDS PAGE analysis 1 μg of recombinant protein was separated by reducing SDS PAGE and visualised by Coomassie Blue staining. Caithness Biotech recombinant LBP migrates at approximately 63 kDa due to glycosylation.		Figure 2: Validation of low levels of TLR2 and/or TLR4 stimulating contaminants HEK-293 cells were transfected with NF-κB reporter and CD14 together with TLR2 or TLR4 and MD2, then treated with indicated concentrations of Pam3CSK4, or E. coli LPS, or the reconstituted protein at 1 μg/ml. NF-κB signalling was measured after overnight treatment by luminometry.
		Product flyer Please click here to download the product flyer.		Technical data sheet Please click here to download the product technical data sheet.
	References
	[1] Gutsmann T, Müller M, Carroll SF, MacKenzie RC, Wiese A, Seydel U. Dual role of lipopolysaccharide (LPS)-binding protein in neutralization of LPS and enhancement of LPS-induced activation of mononuclear cells. Infect Immun 69:6942-50 (2001) [2] Thompson PA, Tobias PS, Viriyakosol S, Kirkland TN, Kitchens RL. Lipopolysaccharide (LPS)-binding protein inhibits responses to cell-bound LPS. J Biol Chem 278:28367-71 (2003) [3] Han Y-H, Onufer EJ, Huang L-H, Sprung RW, Davidson WS, Czepielewski RS, Wohltmann M, Sorci-Thomas MG, Warner BW, Randolph GJ. Enterically derived high-density lipoprotein restrains liver injury through the portal vein. Science 373:eabe6729 (2021)

 

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Caithness Biotechnologies Ltd., 72 Boston Road, Leicester, UK, LE4 1HB. Telephone: +44 (0) 116 326 3802 | email: contact@caithnessbiotechnologies.com