Chicken antibodies isolated from egg yolks (IgY) are a suitable alternative to conventional antibodies from blood of experimental animals. Main advantages of IgY are: (i) larger amounts of IgY could be prepared from repeatedly laid chicken eggs, (ii) collection of eggs is a non-invasive procedure contrary to bleeding of mammals, and (iii) a better response of chicken to mammalian antigens due to the larger evolutionary distance. Here, we report a cost- and labor-effective two-step procedure consisting of yolk extraction by tap water (8-fold dilution, freezing, and filtration) followed by a specific precipitation of IgY at pH 4 with sodium chloride in the final concentration of 8.8%. Using this procedure, the highly purified antibody (97%) was prepared. Resulting IgY preparations are acceptable for a wide range of applications as it was prepared using only bio-compatible chemicals. The purification protocol was developed and optimized in terms of the time, materials and the necessary manipulations.