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LOCALIZATION OF MYOGLOBIN IN HUMAN SKELETAL MUSCLE USING FLUORESCENT ANTIBODY TECHNIQUE

LAWRENCE J. KAGEN 1 and RAYA GUREVICH 1

1 Department of Medicine, Columbia University College of Physicians and Surgeons, and the Edward Daniels Faulkner Arthritis Clinic of the Presbyterian Hospital, New York, New York

Rabbit antiserum to human myoglobin was used with the indirect fluorescent antibody technique to localize this protein in human skeletal muscle. Specific fluorescence was noted, in rapidly frozen and acetone-fixed sections, to be located at the transverse striations, at the sarcolemmal regions and at certain fibrillar structures within the cell. The antibody fluorescence reaction was shown to be specific for myoglobin, and was not produced by normal rabbit serum of serum of rabbits immunized with bacterial antigens. The reaction was abolished by prior absorption of the antimyoglobin serum with myoglobin, and was found to be absent in tissues deficient in myoglobin (lung, kidney, spleen, liver and uterus). Omission of acetone fixation or delayed freezing resulted in leakage of myoglobin from the cell and loss of specific intracellular localization. Sarcolemmal localization appeared to be somewhat more stable.

Submitted on March 2, 1967


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