The ImmunoDiangostics Human NGAL Immunoassay is a Solid Phase ELISA that is designed to accurately detect NGAL in human serum, plasma, urine, and cell culture supernatant samples.
The ImmunoDiagnostics Lipocalin-2 (NGAL) ELISA is a reliable and reproducible tool for the quantitative detection of NGAL antigen. The assay is intended for use on human serum, plasma, urine, or cell culture supernatants.
Numerous application fields exist, but particularly:
- Nephrology: Reports suggest that NGAL may represent a sensitive biomarker for various renal injuries.
- Oncology: NGAL has been shown to be upregulated in a variety of different cancers cells including: breast, ovarian, colorectal, and pancreatic.
- Metabolic Syndrome: A growing body of research suggests that serum levels of NGAL correlate to obesity, insulin resistance, and fatty liver disease (among others).
NGAL Antibody Coated Microstrips: One microplate, 12 strips with 8 wells each, 96 dry wells in total. The wells are coated with a monoclonal anti- human NGAL antibody. The microplate is sealed in a foil bag. Any unused strips should be returned to the bag and resealed for future use.
Detection Antibody Solution: Concentrate (100x conc). One vial containing 0.12 mL of an HRP conjugated mouse monoclonal anti- human NGAL. Detection antibody should be diluted with only the 1X Assay Buffer needed.
Assay Buffer: Concentrate (5x conc). One vial containing 20 mL of buffer for dilution of the detection antibody. If precipitates are observed in the concentrated buffer, warm at 37oC until the precipitates disappear. Dilute 20 mL of 5x Assay Buffer with 80 mL diH2O to make a 1x concentration prior to use.
Human NGAL Standard: 25 ng of recombinant human NGAL in a buffered protein base, lyophilized. Reconstitute with 1 mL 1x Assay Buffer to prepare 25 ng/mL standard stock. Prepare a serial dilution series with 1x Assay Buffer. The reconstituted standard stock can be aliquoted and stored at -20oC for one month. Avoid repeated freeze/thaw cycles.
Wash Buffer: Concentrate (10x conc). One vial containing 50 mL of 10x wash buffer. If precipitates are observed in the 10x solution, warm at 37oC until the precipitates disappear. Prepare 1x Wash Buffer my mixing 50 mL of 10x Wash Buffer with 450 mL diH2O.
Substrate Solution: One bottle containing 12 mL of TMB (3,3’,5,5’-Tetramethylbenzidine) substrate. Ready-to-use. Do not expose to light.
Stop Solution: One vial containing 12 mL of ready-to-use stop solution.
The NGAL ELISA is a quantitative sandwich enzyme immunoassay. Standard, samples, and any desired controls are pipetted into the wells pre-coated with solid-phase capture antibody. After washing away any unbound substances, an HRP-labelled detection antibody, specific for human NGAL, is added to the wells, and any unbound antibody is washed out. A TMB solution is added to develop color in proportion to the amount of human NGAL initially bound. The color development is stopped by addition of an acidifying solution and the optical density is determined. By plotting a standard curve versus measured absorbance, the amount of antigen in the sample can be calculated. The concentration of the antigen is expressed in ng/mL.
The kit contains calibrators and can be used in any laboratory equipped with a plate reader. The kit is optimized for the quantitative determination of human NGAL concentrations in serum plasma, and cell culture supernatant samples. The test can provide results within 3 hours.
Example only! Do not use for calculations
4 Parameter Curve
Log-log Regression Curve
Lipocalin-2, also called neutrophil gelatinase-associated lipocalin (NGAL), is a secretory glycoprotein expressed in liver, lung, kidney, adipocytes, activated neutrophils, and macrophages and can be used as a biomarker for kidney injury.
The physiologic role of NGAL is in innate immunity, binding to bacterial siderophores and sequestering the iron containing compounds to prevent their use by the invading pathogen. NGAL also functions as a growth factor.
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