Assay NGAL (human neutrophil gelatinase-associated lipocalin) levels in human serum, plasma (EDTA, heparin, sodium citrate), tissue extracts, urine and culture supernatant samples with our Thermo Scientific Human NGAL ELISA Kit.
This human NGAL ELISA kit uses pre-titered, matched pairs of coating and detection antibodies to achieve sensitive and accurate measurement of neutrophil gelatinase-associated lipocalin (NGAL). The assay is a sandwich ELISA performed in microplate wells coated with a monoclonal antibody against human NGAL. Bound NGAL is detected with another monoclonal antibody labeled with biotin and the assay is developed with horseradish peroxidase (HRP)-conjugated streptavidin and a color-forming substrate. The assay is a four-step procedure.
- Target: natural and recombinant human NGAL (human neutrophil gelatinase-associated lipocalin)
- Format: colorimetric ELISA (TMB substrate) in clear 96-well strip plate
- Measurement: absorbance at 450nm minus absorbance at 550nm
- Assay Range: 4 to 1000pg/mL
- Sensitivity: < 2pg/mL
- Precision: undetermined
- Sample Types: cell culture supernatant, tissue extracts, serum, plasma (EDTA, sodium citrate and heparin) and urine
- Total Assay Time: 3.5 hours
- Sample Size: 100μL of 10-fold diluted sample per well
- Specificity: The two monoclonal antibodies against human NGAL used in the assay have been shown to bind to different preparations of recombinant human NGAL and to give a single band at 25kDa on Western blot analysis of a reduced postnuclear supernatant from human neutrophils.
Complete kit contains pre-coated antibody-plate, detection antibodies, buffer, diluent, standards, and substrate.
NGAL (neutrophil gelatinase-associated lipocalin) belongs to the lipocalin family of proteins. These are typically small secreted proteins characterized by their ability to bind small, hydrophobic molecules in a structurally conserved pocket formed by beta-pleated sheet, to bind to specific cell-surface receptors and to form macromolecular complexes. NGAL has many synonyms: it also known as NL (neutrophil lipocalin; HNL: human NL), lipocalin 2, oncogene protein 24p3 or uterocalin (in the mouse) and neu-related lipocalin or 25kDa alpha-2-microglobulin-related protein (in the rat). Human NGAL consists of a single disulfidebridged polypeptide chain of 178 amino-acid residues with a calculated molecular mass of 22kDa, but glycosylation increases its apparent molecular mass to 25 kDa. In neutrophils (neutrophilic polymorphonuclear leukocytes) and urine it occurs as monomer, with a small percentage of dimer and trimer, and also in complex with 92-kDa human neutrophil type IV collagenase, also called gelatinase B or matrix metalloproteinase-9 (MMP-9). NGAL is often used as a marker of acute renal injury which may lead to acute renal failure.