Immunoblotting studies with antisera against Z-protein, desmin, and alpha-actinin showed that Z-protein is clearly distinguishable from desmin and alpha-actinin. Z-protein is not a proteolytic product of another protein but is an intrinsic component of chicken breast muscle myofibrils. In these experiments, an SDS extract of intact muscle was first electrophoresed in a polyacrylamide gel, and then proteins were transferred to a nitrocellulose paper sheet. Detection of each protein on the sheet was made possible by the application of the indirect immunofluorescence technique with the respective antiserum. Immunofluorescence microscope studies using these antisera revealed that Z-protein has the same distribution as alpha-actinin in isolated Z-disk sheets. Anti-Z-protein antiserum and anti-alpha-actinin antiserum stained the interior of Z-disks. On the other hand, antiserum against desmin stained the periphery of Z-disks in isolated Z-disk sheets.

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