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Fig. 5. RACK-I and PKC-ß are complexed on melanosomes. (A) Purified melanosomes were subjected to immunoprecipitation using a monoclonal antibody against RACK-I. Immunoprecipitated proteins were then separated in 7.5% SDS-PAGE, transferred onto a nitrocellulose membrane, and the membrane was then cut at the molecular weight of 50 kD. The upper part of the membrane was immuno-reacted using a monoclonal antibody against PKC-ß and the bottom part of the membrane was immuno-reacted using a monoclonal antibody specific for RACK-I. (B) Purified melanosomes were subjected to immunoprecipitation using a monoclonal antibody against RACK-I. Then the immunoprecipitated proteins were separated and immunoblot analysis was performed using specific antibody against PKC-{alpha} on the upper part of the membrane and monoclonal antibody against RACK-I. Purified recombinant PKC-{alpha} (5 ng) purchased from Panvera was used as a control.





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