β-Casomorphin, human

To study the effect of β-Casomorphin, human and DAMGO, cells are seeded in 12-well culture plates. Experiments are performed 3 (DHE cells) or 21 days (HT29-MTX cells) after confluency. Twenty-four hours before the studies, the culture medium is replaced by serum-free medium to starve the cells from serum and to eliminate any interference from extraneous proteins or hormones. The experimental protocol is then the following: the serum-free medium is removed, and the monolayer cultures of DHE or HT29-MTX are washed twice with PBS (37°C). Serum-free medium with or without β-Casomorphin, human or DAMGO is added to the cells and incubated at 37°C for 30 min to 24 h in a humidified atmosphere. μ-Opioid receptor blockade is performed by preincubating the cells with cyprodime for 30 min before agonist addition. The supernatants are then collected, frozen, and stored at 20°C. Cells are processed with trypsin. The cell numbers per well are determined, and total RNA is isolated. All experiments are performed at least three times in triplicate^[2].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.