Description

Nori Human Galectin-3 ELISA Kit Summary

Alternative names for Galectin-3: Galectin 3, LGALS3

Standard Curve

Kit Components
1. Pre-coated 96-well Microplate
2. Biotinylated Detection Antibody
3. Streptavidin-HRP Conjugate
4. Lyophilized Standards
5. TMB One-Step Substrate
6. Stop Solution
7. 20 x PBS
8. Assay Buffer

Other Materials Required but not Provided:
1. Microplate Reader capable of measuring absorption at 450 nm
2. Log-log graph paper or computer and software for ELISA data analysis
3. Precision pipettes (1-1000 µl)
4. Multi-channel pipettes (300 µl)
5. Distilled or deionized water

Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the datasheet.
2. Add 100 µl of Standard or samples to each well and incubate 1 h at RT.
3. Add 100 µl of Working Detection Antibody to each well and incubate 1 h at RT.
4. Add 100 µl of Working Streptavidin-HRP to each well and incubate 20 min at RT.
5. Add 100 µl of Substrate to each well and incubate 5-30 min at RT.
6. Add 50 µl of Stop Solution to each well and read at 450 nm immediately.

Background: 

Galectin-3 is a protein that in humans is encoded by the LGALS3 gene.^[1] Galectin-3 is a member of the lectin family, of which 14 mammalian galectins have been identified. Galectin-3 is approximately 30 kDa and, like all galectins, contains a carbohydrate-recognition-binding domain of about 130 amino acids that enable the specific binding of β-galactosides.^[3] Galectin-3 (Gal-3) is also a member of the beta-galactoside-binding protein family that plays an important role in cell-cell adhesion, cell-matrix interactions, macrophage activation, angiogenesis, metastasis, apoptosis. Galectin-3 is expressed in the nucleus, cytoplasm, mitochondrion, cell surface, and extracellular space.^[2] Galectin-3 has an affinity for beta-galactosides and exhibits antimicrobial activity against bacteria and fungi. Given galectin-3’s broad biological functionality, it has been demonstrated to be involved in cancer, inflammation and fibrosis, heart disease, and stroke.^[3] Studies have also shown that the expression of galectin-3 is implicated in a variety of processes associated with heart failure, including myofibroblast proliferation, fibrogenesis, tissue repair, inflammation, and ventricular remodeling.^[4] Galectin-3 associates with the primary cilium and modulates renal cyst growth in congenital polycystic kidney disease.^[5] Chronic heart failure has been found to be indicated by a galectin-3 tests, the myocardial lining constantly undergoes necessary fibrosis, and the inhibition of galectin-3 interferes with myocardial fibrogenesis. A study concluded that drugs binding to galectin-3 will benefit those who have too much fibrosis on the heart, but it might potentially backfire for those who need heart restructuring.^[6] Galectin-3 is upregulated in patients with idiopathic pulmonary fibrosis. The cells that receive galectin-3 stimulation (fibroblasts, epithelial cells, and myofibroblasts) upregulated the formation of fibrosis and collagen formation.^[6] Galectin-3 is increasingly being used as a diagnostic marker for different cancers and has varying effects in different types of cancer.^[7]

References

1. Raz A, et al. (1991). Cancer Research. 51 (8): 2173–8.
2. Liu FT, et al. (2002). Biochimica et Biophysica Acta. 1572 (2-3): 263–73.
3. Sharma UC, et al. (2004). Circulation. 110 (19): 3121–8.
4. Lin YH, et al. (2009). Clinica Chimica Acta; Int J Clinical Chem. 409 (1-2): 96–9.
5. Chiu MG, et al. (2006). The American Journal of Pathology. 169 (6): 1925–38.
6. Yu L, et al. (2013) Circulation: Heart Failure. 6 (1): 107–17.
7. Idikio HA (2011). PLoS One. 6 (10): e26150.

Product Citations