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ULTRASTRUCTURAL AUTORADIOGRAPHIC DETECTION OF INTRACELLULAR IMMUNOGLOBULINS WITH IODINATED Fab FRAGMENTS OF ANTIBODY THE COMBINED USE OF ULTRASTRUCTURAL AUTORADIOGRAPHY AND PEROXIDASE CYTOCHEMISTRY FOR THE DETECTION OF TWO ANTIGENS (DOUBLE LABELING)

NICHOLAS K. GONATAS 1, ANNA STIEBER 1, JACQUELINE GONATAS 1, PIERLUIGI GAMBETTI 1, JEAN CLAUDE ANTOINE 2, and STRATIS AVRAMEAS 2

1 Department of Pathology, Division of Neuropathology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104
2 The Laboratory of Immunocytochemistry, Pasteur Institute, Paris, France

Intracellular immunoglobulins of paraformaldehyde-fixed rat lymph node cells were identified with ultrastructural autoradiography of 125I Fab of sheep antirat Ig antibody. We observed excellent intracellular penetration and specific localization of the iodinated Fab fragments of the antirat Ig antibody. In contrast, 125I sheep antirat IgG or conjugates of peroxidase and Fab-antibody fragments did not show satisfactory intracellular penetration. Simultaneous detection of plasma membrane and intracellular immunoglobulins has been achieved with the use of iodinated Fab of sheep antirat Ig and peroxidase-labeled Fab of sheep antirat Ig and subsequent processing of the cells for cytochemical (peroxidase) and autoradiographic study. The simultaneous demonstration of specific (antiperoxidase) antibody and Ig molecules is also feasible.

Submitted on May 2, 1974
Revised on July 23, 1974


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C Lin and J Chang
Electron microscopy of albumin synthesis
Science, October 31, 1975; 190(4213): 465 - 467.
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