Method for simultaneous isolation of myoglobin, cytochrome c and hemoglobin from tissue

A method was developed for the simultaneous isolation and purification of myoglobin, hemoglobin, and cytochrome c from muscle tissue. Two standard methods for the individual isolation of myoglobin and hemoglobin and of cytochrome c were modified to achieve high yields of the chromoproteins from the same tissue specimen. The relative yields of myoglobin and hemoglobin by this modified method are somewhat higher than in the original method. The recovery of cytochrome c utilizing the modified method was approximately 83% when compared with the original methodology. The original methods for deriving cytochrome c and myoglobin from tissues would normally require the use of two different tissue samples. The primary advantage of this combined method is therefore economy of tissue utilized in the recovery of these chromoproteins. In many applications the availability of tissue is very often limited by the small size of the experimental animal, the rarity of the species, or both. Secondarily the combined method affords true comparative data since pigments are actually isolated for the same cellular population.