Igg streptavidin biotin complex kit

Human lung and mouse lung tissues were stained with rabbit monoclonal MBD2 antibody (1:500, Abcam, Cambridge, UK) and rabbit polyclonal MBD2 antibody (1:100, Santa Cruz Biotechnology, CA, USA), respectively. These proteins were then detected in lung sections with an IgG Streptavidin Biotin Complex kit (Boster, Wuhan, People’s Republic of China) and developed with DAB substrate according to the manufacturer’s protocol (Dako, Glostrup, Denmark). A Nikon Spot image acquisition and processing system (USA) was used for image assessment. The mean integral OD of MBD2 protein staining in the bronchiolar epithelium was measured as follows.32 (link) We used Image-Pro Plus 4.1 professional image analysis software to open all immunohistochemical images from one sample slide. Thereafter, we activated the Magnetic Lasso tool and dragged selection areas only containing bronchial epithelium, then copy and pasted it into a new image. Then, the area of bronchial epithelium and the integral OD of MBD2-positive epithelium were measured. Finally, the mean integral OD of MBD2 protein staining in the bronchial epithelium was the ratio of the integral OD of MBD2-positive epithelium to area of bronchial epithelium. All of the images are assessed with the same parameters.