Mouse anti cardiac troponin t ctnt

Biowires were fixed with 4% paraformaldehyde, permeablized by 0.25% Triton X-100, and blocked by 10% bovine serum albumin (BSA). Immunostaining was performed using the following antibodies: mouse anti-cardiac Troponin T (cTnT) (Abcam; 1:100), rabbit anti-Connexin 43 (Cx-43) (Abcam; 1:200), mouse anti-α-actinin (Abcam; 1:200), goat anti-mouse-Alexa Fluor 488 (Jackson Immuno Research; 1:400), anti-rabbit-TRITC (Invitrogen; 1:200), anti-mouse-TRITC (Jackson Immuno Research; 1:200). Nuclei were counterstained with 4',6-diamidino-2-phenylindole (DAPI) (Biotium; 1:100). Phalloidin-Alexa 660 (Introgen; 1:600) was used to stain F-actin fibers. For confocal microscopy, the stained cardiac biowires were visualized under an inverted confocal microscope (Olympus IX81) or an upright confocal microscope (Zeiss LSM 510).

At the final time point of six days, cardiac microtissues were fixed in 4% paraformaldehyde. Immunostaining was performed as described previously [5 (link)] using the following antibodies: mouse anti-cardiac troponin T (cTNT) (Abcam; 1:100), mouse anti-α-actinin (Abcam, 1:20), rabbit anti-connexin 43 (Cx43) (Abcam, 1:500), goat anti-mouse-Alexa Fluor 488 (Jackson ImmunoResearch; 1:400) and donkey anti-rabbit-TRITC (Invitrogen; 1:400). Phalloidin (Invitrogen 1:100) was used to detect actin fibers. Cells were visualized using a fluorescence confocal microscope (Zeiss LSM-510).