Galectin 3 human simplestep elisa kit

For the assay, cells were seeded at 1.8 × 10⁵ cells/1 ml medium/3.8 cm² growth area in 12-well plates, grown overnight, and treated with morin and/or cisplatin for 24 h and/or 48 h. Concentrations of drugs’ solvents were corrected in all wells (including control wells) to the constant level, corresponding to the highest used concentration of a particular solvent. Total protein extraction and ELISA assay were performed using Galectin-3 Human SimpleStep ELISA^® Kit (Abcam) and the concentrations of proteins in solutions were determined using Bradford Reagent (Sigma-Aldrich) and Bovine Serum Albumin Standard Set (Fermentas) according to the manufacturers’ protocols. The absorbance of each well was measured at 450 nm (ELISA assay) or 595 nm (Bradford assay) with ELISA plate reader (Dynex Technologies Triad Multi-Mode Microplate Reader). The concentration of galectin-3 in 1 µg of protein extract in each sample was determined by interpolating the blank control subtracted absorbance values against the standard curve and by multiplying the resulting value by the appropriate sample dilution factor. Samples were prepared in triplicate and absorbance measurement for each sample was done in duplicate.

Total protein extraction from the cells reverse transfected with RNA oligonucleotides in 24-well plates (24 h, 48 h, and 72 h after reverse transfection) followed by ELISA assay was performed using Galectin-3 Human SimpleStep ELISA^® Kit (Abcam) according to the instruction manual. The protein concentration of the samples was determined using Bradford Reagent (Sigma-Aldrich), according to the manufacturer’s protocol, supplemented with Bovine Serum Albumin Standard Set (Fermentas). The absorbance of each sample was measured with ELISA plate reader (Dynex Technologies Triad Multi-Mode Microplate Reader) at 595 nm (Bradford assay) or 450 nm (ELISA assay). In each sample, the concentration of galectin-3 in 1 µg of total protein extract was calculated by interpolating the blank control subtracted absorbance values against the standard curve, followed by multiplying the resulting value by dilution factor. Samples were prepared in triplicate and absorbance measurement for each sample was done in duplicate.