Detection Of β-Casein In Bovine Mammary Epithelial Cells

The purpose of this article is to establish methods which can detect β-casein of cultured bovine mammary epithelial cells in vitro exactly and conveniently. These methods can be used to measure the physiological function and differentiation of cultured bovine mammary epithelial cells in vitro. And it is also useful to estimate expression of casein regulated by hormones at cell level. Established the methods to detect β-casein at protein level and transcription level. First, we used the bovineβ-casein purchased from Sigma company to immunize the New Zealand Rabbits. And we obtained the antibody of β-casein.At protein level, we established the two ELISA methods to detectβ-casein using the obtained antibody. And we compared the double antibody Sandwich ELISA with indirect competitive ELISA. The result of experiment indicated that the sensitivity of double antibody Sandwich ELISA was better than indirect competitive ELISA.At transcription level, we detected transcription of β-casein using RT-PCR method. Using these two methods we detected theβ-casein in mammary epithelial cells separated from beastings.The expression ofβ-casein must be induced by some hormones. In this experiment, we attempted to use Prolactin ,Insulin and Hydrocordisone to induce the cultured mammary epithelial cells in vitro. And we detected theβ-casein using double antibody ELISA and RT-PCR methods . But we didn't found theβ-casein in it.