Connexin 43 monoclonal antibodies

Immunofluorescence staining was performed in cells fixed with paraformaldehyde. The fixed cells were washed with PBS and then incubated with 2% goat serum and 5% bovine-serum albumin in PBS to reduce nonspecific binding. The cells or cardiac sections were then incubated for 2 hours at room temperature with mouse, anti-α-sarcomeric actinin (1:500, Sigma-Aldrich, MO), and connexin-43 monoclonal antibodies (1:500, Cell Signaling, MA). The sections were then incubated with the appropriate goat or rabbit anti-mouse secondary antibodies (1:1000) conjugated to Texas red and FITC. The nuclei were counterstained with HardSet mounting medium with DAPI (Vector Labs). The cells were visualized by Olympus FV1000 spectral confocal microscopy. Separate cells were also stained without primary antibodies to identify nonspecific binding.

Immunofluorescence staining was performed in cells or cardiac sections fixed with paraformaldehyde. The fixed cells were washed with PBS and then incubated with 2% goat serum and 5% bovine-serum albumin in PBS to reduce nonspecific binding. The cells or cardiac sections were then incubated for 2 hours at room temperature with mouse, anti-α-sarcomeric actinin (1∶500, Sigma-Aldrich, MO), cardiac troponin-T (1∶200, Santa Cruz, CA), myosin heavy chain (1∶200, Santa Cruz, CA) and connexin-43 monoclonal antibodies (1∶500, Cell Signaling, MA). The sections were then incubated with the appropriate anti-mouse secondary antibodies (1∶1000) conjugated to Texas red and FITC. The nuclei were counterstained with HardSet mounting medium with DAPI (Vector Labs). The cells were visualized by inverted Nikon fluorescence microscope (TE 2000). Separate cells were also stained without primary antibodies to identify nonspecific binding.