Immunofluorescent staining of frozen mouse retina using Zymed Rb anti-Connexin 57 (Mid) (Cat. No. 40-5000). Image courtesy of James I. Nagy, PhD, University of Manitoba, Canada.
Immunohistochemistry analysis of STAT4 showing staining in the cytoplasm of paraffin-embedded human heart tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a STAT4 monoclonal antibody (405000) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.