The ImmunoDiangostics Human Galectin-3 Immunoassay is a Solid Phase ELISA that is designed to accurately detect Galectin-3 in human serum and plasma samples.
The ImmunoDiagnostics Galectin-3 (ELISA) is a reliable and reproducible tool for the quantitative detection of Galectin-3 antigen. The assay is intended for use on human serum or plasma.
Numerous application fields exist, but particularly:
- Hepatology: The expression of Galectin-3 is induced in cirrhotic liver and hepatocellular carcinoma while its ablation protects mice from developing diet-induced NASH.
- Cardiovascular Disease: Galectin has been linked to tissue repair, cardiac remodeling, and proliferation of myofibroblasts in heart failure and can be a useful aid in the prognosis of heart failure outcome.
Oncology: Altered expression of Galactin-3 can affect cancer cell growth and differentiation, apoptosis, angiogenesis, as well as invasion and metastasis. Additionally, is can be used as a prognostic factor for predicting the progression of cancer.
Galectin-3 Antibody Coated Microstrips: One microplate, 12 strips with 8 wells each, 96 dry wells in total. The wells are coated with a polyclonal anti- human Galectin-3 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 polyclonal anti- human Galectin-3. 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 Galectin-3 Standard: 40 ng of native human Galecin-3 in a buffered protein base, lyophilized. Reconstitute with 1 mL 1x Assay Buffer to prepare 40 ng/mL standard stock. Prepare a serial dilution series with 1x Assay Buffer. The reconstituted standard stock can be aliquoted and stored at -80oC 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 Galectin-3 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, a biotin-labelled detection antibody, specific for human Galectin-3, is added to the wells, and any unbound antibody is washed out. Streptavidin-HRP conjugate is then added and a TMB solution is added to develop color in proportion to the amount of human Galectin-3 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 Galectin-3 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
Galectin-3 is a member of the lectin family of proteins. More specifically, galectin has strong affinity for β-galactosidase and has been shown to exhibit antimicrobial activities. Physiologically, galectin-3 is involved with cell growth and differentiation, cell adhesion, chemoattraction and cell activation, the cell cycle, and apoptosis. Unsurprisingly, galectin-3 can be involved with cancer, inflammation and fibrosis, as well as heart disease.
Galectin-3 expression has been correlated to multiple forms of fibrosis: hepatic, renal, and idiopathic pulmonary fibrosis. Its expression is further induced in subjects with NASH, cirrhosis, and hepatocellular carcinoma.
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