The NGAL (Neutrophil Gelatinase-Associated Lipocalin) ELISA Kit is designed for the quantitative measurement of NGAL levels in various biological samples, such as serum, plasma, urine, and tissue homogenates. NGAL, also known as Lipocalin-2, is a protein that plays a critical role in inflammation and is a biomarker for kidney injury, infection, and other conditions.
Content of the Kit
- Pre-coated 96-Well Microplate:
- Wells pre-coated with an antibody specific to NGAL, allowing capture of NGAL from samples.
- Standards:
- Solutions containing known concentrations of recombinant or purified NGAL, used to generate a standard curve for quantifying NGAL levels in 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 NGAL, 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 NGAL.
Assay Procedure Summary
- Sample and Standard Addition: Add standards and samples to the microplate wells where NGAL is captured by the immobilized antibody.
- Wash: Remove unbound substances by washing the wells.
- Detection Antibody Addition: Add a detection antibody specific to NGAL, which binds to the captured NGAL.
- Wash: Wash away excess detection antibody.
- Enzyme Conjugate Addition: If using a biotinylated detection antibody, add a streptavidin-HRP conjugate to bind to the biotinylated antibody.
- Wash: Remove unbound enzyme conjugate through washing.
Performance Characteristics
- Sensitivity: Capable of detecting low levels of NGAL, typically 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.
- Recovery: High recovery rates, usually above 85%, indicating reliable performance across various sample types.
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