| Field | Specification |
|---|---|
| Target | |
| Alternative names | Lipopolysaccharide-binding protein;LBP;Lbp; |
| Gene ID | |
| UniProt # | |
| Reactivity | |
| Applications | |
| Immunogen | Expression system for standard: NS0; Immunogen sequence: G25-V481 |
| Expression system | |
| Expression region | |
| Assay type | |
| Sample type(s) | Cell Culture Supernatant, Cell Lysate, Serum, Plasma |
| Sensitivity | |
| Detection range | |
| Assay time | |
| Storage | |
| Catalog no. (Mfr.) | |
| Main SKU |
Assay Principle
This sandwich ELISA quantifies mouse lipopolysaccharide-binding protein (gene Lbp) in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Strips are pre-coated with a capture antibody. After the sample is added, a biotinylated detection antibody binds the captured protein, and an avidin–biotin–peroxidase complex supplies the enzyme. TMB develops color in proportion to the bound target; the reaction is stopped and read at 450 nm, and concentrations are read from the standard curve.
Lipopolysaccharide-binding protein (LBP) is an acute-phase plasma protein made mainly by the liver. It binds the lipid A part of bacterial lipopolysaccharide and hands it to CD14, which then presents it to the TLR4/MD-2 receptor, greatly increasing cell sensitivity to Gram-negative bacteria. LBP is studied in innate immunity, sepsis and inflammation research.
Kit Components
| Anti-Mouse Lbp Pre-coated 96-well strip microplate | 1 plate (96 wells) |
|---|---|
| Mouse Lbp Standard | 2 vials, 100 ng/tube |
| Mouse Lbp Biotinylated antibody (100x) | 100 µL |
| Avidin-Biotin-Peroxidase Complex (100x) | 100 µL |
| Sample Diluent | 30 mL |
| Antibody Diluent | 12 mL |
| Avidin-Biotin-Peroxidase Diluent | 12 mL |
| Color Developing Reagent (TMB) | 10 mL |
| Stop Solution | 10 mL |
| Wash Buffer (25x) | 20 mL |
| Adhesive plate sealers | 4 |
Also required (not supplied): microplate reader for 450 nm; incubator; automated plate washer (optional); precision pipettes for 0.5 µL through 1 mL volumes of aqueous solutions; multichannel pipettes (recommended for many samples); deionized or distilled water; 500 mL graduated cylinders; test tubes for dilution.
Kit components are not sold separately.
Performance Data
| Species | Mouse |
|---|---|
| Sample types | Cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA) |
| Assay range | 1.56 ng/mL–100 ng/mL |
| Sensitivity | <50 pg/mL |
| Standard | Recombinant mouse Lbp (Gly25–Val481), expressed in NS0 |
| Cross-reactivity | None detected against related proteins in supplier testing |
| Intra-assay CV | 5.4–7.1% (3 samples, n = 16 each) |
| Inter-assay CV | 7.1–8.1% (3 samples, n = 24 each) |
| Format | 96 wells, removable strips |
Sample dilution: start at 1:100. In the supplier’s tests, serum and plasma samples measured about 500 ng/mL.
Example standard curve (OD450, ng/mL): 0 = 0.01; 1.56 = 0.073; 3.12 = 0.138; 6.25 = 0.243; 12.5 = 0.484; 25 = 0.907; 50 = 1.475; 100 = 2.004. Run a new standard curve on every plate.
Storage & Handling
Store at 4 °C for up to 6 months or −20 °C for up to 12 months. Avoid repeated freeze–thaw cycles. Ships on gel ice and tolerates up to 3 days at room temperature; freeze on arrival.
Safety
Sample data
| Concentration (ng/ml) | 0 | 1.56 | 3.12 | 6.25 | 12.5 | 25 | 50 | 100 |
| O.D. | 0.01 | 0.073 | 0.138 | 0.243 | 0.484 | 0.907 | 1.475 | 2.004 |
Intra/inter assay consistency
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 16 | 16 | 16 | 24 | 24 | 24 |
| Mean (pg/ml) | 3313 | 12997 | 39089 | 3385 | 14212 | 39495 |
| Standard deviation | 237.12 | 831.8 | 2110.8 | 243.72 | 1151.17 | 2804.14 |
| CV (%) | 7.1% | 6.4% | 5.4% | 7.2% | 8.1% | 7.1% |
Kit components
Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x)|100ul Avidin-Biotin-Peroxidase Complex (100x)|100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB)|10ml Stop Solution|10ml Wash Buffer (25x)|20ml Adhesive plate sealers|4Materials required but not provided
- Microplate Reader capable of reading absorbance at 450nm.
- Incubator.
- Automated plate washer (optional).
- Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
- Multichannel pipettes are recommended for large amount of samples.
- Deionized or distilled water.
- 500ml graduated cylinders.
- Test tubes for dilution.
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