Manual
Description
Lipopolysacharide (LPS) Binding Protein (LBP) is a type 1 acute phase protein that is constitutively produced by the liver and rapidly upregulated during the acute phase response. LBP plays a central role in the response to LPS. The protein catalyses the monomerization of LPS and its transfer to (s)CD14 and to lipoproteins. This way LBP has both a role in the activation pathway of LPS: activation of monocytes by LPS leading to release of inflammatory mediators and in the neutralization of LPS i.e. the uptake of LPS by lipoprotein and subsequent clearing. In plasma of healthy individuals LBP is present at levels of approximately 10 μg/ml, which increase approximately 10-fold during acute phase responses.
Application
The human LBP ELISA has been developed for the quantitative measurement of natural and recombinant human LBP in cell culture medium, plasma or serum. In serum or plasma samples human LBP can be measured accurately if serum or plasma samples are diluted at least 1000 times. Most reliable results are obtained if EDTA plasma is used.
Features
- Minimum concentration which can be measured is 1 ng/ml human LBP.
- Measurable concentration range of 4.4-50 ng/ml.
- Working volume of 100 µl/well.
Typical standard curve
Principle
- The human LBP ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 3½ hours.
- The efficient format of a plate with twelve disposable 8-well strips allows free choice of batch size for the assay.
- Samples and standards are captured by a solid bound specific antibody.
- Biotinylated tracer antibody will bind to captured human LBP.
- Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody.
- Streptavidin-peroxidase conjugate will react with the substrate, tetramethylbenzidine (TMB).
- The enzyme reaction is stopped by the addition of citric acid.
- The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the human LBP standards (log).
- The human LBP concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.
Storage and stability
Product should be stored at 4°C. Under recommended storage conditions, product is stable for at least six months. After reconstitution the reagents are stable for 1 month if stored at 2-8°C. After reconstitution the standard is stable for 24 hours. For longer stability we recommend to store aliquots at -20°C.
References
- Kaser, A et al; Endotoxin and its binding proteins in chronic liver disease: the effect of transjugular intrahepatic portosystemic shunting. Liver 2002, 22: 380
- Labéta, M et al; Innate recognition of bacteria in human milk is mediated by a milk-derived highly expressed pattern recognition receptor, soluble CD14. J Exp Med 2000, 191: 1807
- Ahrén, I et al; Lipopolysaccharide-binding protein increases toll-like receptor 4-dependent activation by nontypeable Haemophilus influenzae. J Infect Dis 2001, 184 : 926
- Lepper, P et al; Association of Lipopolysaccharide-binding protein and coronary artery disease in men. J of Am Coll Cardiol 2007, 50:25
- Gubern, C et al; Natural antibiotics and insulin sensitivity. The role of bactericidal/Permeability-Increasing protein. Diabetes 2006, 55:216
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 of or derivation of this product.
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