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To prevent non-specific binding, myocytes were blocked with 10% normal donkey serum (Sigma®) in PBS overnight (labeling buffer) [44].
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This was performed by removing all the gap junctions of the chosen myocytes, so that any propagation into or out of this myocyte is blocked.
The residual binding sites were blocked through incubation in TBS containing 0.2% Tween and 5% BSA.
To prevent unspecific binding, membranes were blocked with 4% BSA or 1% PVP-T.
Nonspecific binding sites were blocked with TBST/3% milk powder.
Nonspecific protein binding sites were blocked with serum-free protein blocking solution.
Non-specific binding sites were blocked with goat serum.
Endogenous peroxidase activity and nonspecific binding sites were blocked.
Nonspecific binding sites were blocked with 1% BSA.
Non-specific binding sites were blocked with Protein Block (DAKO, Glostrup, Denmark) for 10 minutes.
Non-specific binding sites were blocked using Protein Block (DAKO) for 20 min.
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