LBP ELISA Kit (Lipopolysaccharide Binding Protein)

Details for Product LBP ELISA Kit No. ABIN370808, Supplier: Log in to see
  • BPIFD2
  • Bpifd2
  • Ly88
  • LBP
  • lipopolysaccharide binding protein
  • lipopolysaccharide-binding protein
  • LBP
  • Lbp
  • LOC100472839
Mouse (Murine)
Kits with alternative reactivity to:
Method Type
Sandwich ELISA
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Purpose The mouse LBP kit has been developed for the quantitative measurement of natural and recombinant mouse LBP in serum, plasma and culture medium. The mouse LBP kit is a solid phase sandwich Enzyme-Linked-Immunosorbent Assay (ELISA). Monoclonal antibody specific for mouse LBP is used for precoated modules. In the first step the precoated modules will be incubated with the antigen (standard or sample). During this incubation, mouse LBP is captured by solid bound antibody. Unbound material present in the sample will be removed by washing. Now the plate will be incubated with a POD-labelled antibody specific for mouse LBP (second incubation). Revelation step includes TMB as chromogen. The enzyme reaction is stopped by the addition of stopping solution and the absorption at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorptions versus the corresponding concentrations of the known standards. The mouse LBP concentration of samples with unknown concentrations, which are run concurrently with the standards, can be determined from the standard curve.
Analytical Method Quantitative
Detection Method Colorimetric
Specificity Cross reaction: rat LBP. Specificity: detected free as well as bound LBP. Recovery of recombinant LBP in LBP depleted sera is 100%
Sensitivity Normal LBP range in untreated mice: (2-15ug/ml). Acute phase sera containing factor 10 to 100 more LBP. Interassay variation coefficient: 7% till 13.6% depending of concentration. Intraassay variation coefficient: 2.4%, n=50 plasma samples. Effective range: 1 -50 ng/ml
Characteristics Enzyme Immunoassay for Determination of mouse LBP (useful also for rat LBP)
Material not included Orbital shaker, Micro plate reader for measurement absorbance at 450 /620 nm, Precision pipettes with disposable tips, 10-1000 ul adjustable multiwell pipettes
Plasmids, Primers & others Plasmids, Primers & others LBP products on genomics-online (e.g. as negative or positive controls)
Alternative Name Lipopolysaccharide-binding Protein (LBP) (LBP ELISA Kit Abstract)
Pathways TLR Signaling, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Positive Regulation of Immune Effector Process, Toll-Like Receptors Cascades, Monocarboxylic Acid Catabolic Process
Application Notes Preparation of reagents: (A) Wash Buffer: PBS/ Tween 0.05%: Dissolve 1 Tablet Phosphate buffered saline (PBS, vial 5) in 200ml distilled water -add 100 ul Tween 20 ( vial 7). (Prepared wash buffer is stable for 4 weeks at refrigerator). B PBS: Dilute 1 Tablet of vial 5 in 200 ml distilled water. C Dilution buffer: Add content of the vial 6 to 50ml PBS (Buffer C). Prepare just before use. Store remaining dilution buffer after reconstitution at -20oC. D Substrate: Vial 9 Ready for use, mix carefully. E Detection antibody: Vial 2 Ready for use, mix carefully. F mouse reference serum: Add 10 ul distilled water to the vial 4. This contains 10.4
Plate Pre-coated
Protocol Let all reagents reach room temperature and mix thoroughly. 1. Samples: Add 100 ul of standards (50, 25, 12.5, 6.25, 3.12 ng/ml= vial a-f) or diluted samples in duplicate into the corresponding wells of the precoated modules and incubate for one hour at room temperature and shaking. 2. 3 x washing with Wash Buffer (A). 3. Detection antibody: Add 100 ul detecting antibody (E) to each well and incubate at room temperature for 1 hour at shaker. 4. 3 x washing with Wash Buffer (A). 5. Substrate: Add 100 ul Substrate solutions (D, vial 9) to each well. Incubate 12-15 min in the dark at room temperature without shaking. 6. Blocking: Add 100 ul stopping solution (vial 8) to each well. Tape gently to mix plate. 7. Read absorbance at 450 nm (reference wave length 620) 8. Calculate the LBP concentration: Calculate the mean of optical density (OD) of standard duplicates, reference serum and the samples. Design a standard curve by plotting the OD means of standards (b-f) (y-axis) and the LBP concentration (x-axis). Calculate the LBP concentration from the mean OD of samples from the standard curve and multiply with dilution factor.
Restrictions For Research Use only
Storage -20 °C
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Product cited in: Depke, Steil, Domanska, Völker, Schütt, Kiank: "Altered hepatic mRNA expression of immune response and apoptosis-associated genes after acute and chronic psychological stress in mice." in: Molecular immunology, Vol. 46, Issue 15, pp. 3018-28, 2009 (PubMed).

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