Probiotic therapy is effective in the treatment of gastrointestinal disorders. It also has immunomodulatory effects due to producing bacteriocins and Bacteriocin-like components. Cholesterol is an important compound in most of the biological reactions. The excess of cholesterol can be seen as a harmful compound of causing heart diseases. In this research, antimicrobial activity and cholesterol removal of 79 isolates were assessed. These isolates had been isolated from traditional dairy products (such as cheeses, yougurth, dough, butter and whey) and showed probiotic potential. Cholesterol removal from media (final concentration 300µg.ml^-1) was measured by o-phetalaldehyde. Based on cholesterol absorbtion, the isolates were classified into three groups. Fisrt group included the isolates whose cholesterol assimilation was less than 100 µg.ml^-1.  This group included three Lactobacillus brevis, four Lactobacillus casei, two Enterococcus faecalis, one Enterocccus sp and one Lactobacillus sp. Second group showed cholesterol assimilation between 100 to 165 µg.ml^-1.  This group included twelve Lactobacillus casei, thirteen Lactobacillus brevis, fourteen Lactobacillus plantarum, sixteen Enterococcus faecium, three Enterococcus faecalis and three Enterococcus durans. Thirth group showed cholesterol assimilation more than 165 µg.ml^-1 that involved two isolates of Enteococcus facium, three isolates of Lactobacillus plantarum and one isolate of Enterococcus faecalis. In the second step, antimicrobial activity of the isolates was determined by agar well diffusion and disc method against seven pathogens including Listeria monocytogenes, Klebsiella pneumoniae, Staphylococcus aureus,  Escherichia coli, Shigella flexneri, Bacillus sereus and Yersinia enterocolitica. Inhibition zone was measured in mm. The maximom inhibition in the isolates was against Listeria monocytogenes and least was observed against Staphylococcus aureus. Fifteen isolates of Lactobacillus brevis and seventeen isolates of Lactobacillus plantarum showed inhibitory zone between 1- 4.5 mm, fourtheen isolates of Lactobacillus casei the zone between 2- 6.5 mm, five isolates of Enterococcus faecalis the zone between 1-4 mm, nineteen isolates of Enterococcus faecium and four isolates of  Enterococcus durans the zone between 1-6 mm, one isolate of Lactobacillus sp the zone between 1-2 mm and one isolate of Enterococcus sp showed the inhibitory zone between 1-6 mm.