The LPS (Lipopolysaccharide) ELISA Kit is designed for the quantitative detection of lipopolysaccharide levels in various biological samples, such as serum, plasma, cell culture supernatants, and other fluid samples. Lipopolysaccharide is a major component of the outer membrane of Gram-negative bacteria and is a potent endotoxin that can induce a strong immune response. Measuring LPS levels is crucial in research related to sepsis, inflammatory responses, and bacterial infections.
Content of the Kit
- Pre-coated 96-Well Microplate:
- Wells pre-coated with an antibody specific to LPS, allowing capture of LPS from the samples.
- Standards:
- Solutions containing known concentrations of LPS, used to generate a standard curve for quantifying LPS levels in unknown samples.
- Sample Diluent:
- A buffer solution used to dilute biological samples to appropriate concentrations for accurate measurement.
- Detection Antibody:
- An antibody specific to a different epitope of LPS, often conjugated to an enzyme such as HRP for detection.
- Enzyme Conjugate:
- If the detection antibody is biotinylated, a streptavidin-HRP conjugate may be included for signal detection.
- Substrate Solution (TMB):
- Tetramethylbenzidine, which reacts with HRP to produce a colorimetric signal proportional to the amount of LPS.
Assay Procedure Summary
- Sample and Standard Addition: Add standards and samples to the microplate wells where LPS is captured by the immobilized antibody.
- Wash: Remove unbound substances by washing the wells.
- Detection Antibody Addition: Add a detection antibody specific to LPS, which binds to the captured LPS.
- Wash: Wash away excess detection antibody.
- Enzyme Conjugate Addition: If using a biotinylated detection antibody, add streptavidin-HRP conjugate to bind to the biotinylated antibody.
- Wash: Remove unbound enzyme conjugate through washing.
Performance Characteristics
- Sensitivity: The assay is capable of detecting low levels of LPS, often in the picogram per milliliter (pg/mL) range.
- Precision: Shows low intra-assay and inter-assay variability, with coefficients of variation (CV) generally below 10%.
- Linearity: Demonstrates linearity across the standard curve range, ensuring accurate quantification of LPS over a wide concentration range.
- Recovery: High recovery rates, usually above 85%, indicating the assay’s reliability and accuracy across various sample types.
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