Detection of antibodies to small nuclear ribonucleoproteins and small cytoplasmic ribonucleoproteins using unlabeled cell extracts

Authors

  • Mark S. Forman,

    Student
    1. Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut
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  • Mary Nakamura,

    Student
    1. Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut
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  • Tsuneyo Mimori MD, PhD,

    Postdoctoral Fellow
    1. Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut
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  • Carmen Gelpi PhD,

    Visiting Scientist
    1. Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut
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    • Department of Immunology, Hospital de la Santa Creu i Sant Pau, Padre Claret, Barcelona, Spain.

  • John A. Hardin MD

    Associate Professor Of Medicine, Corresponding author
    1. Department of Internal Medicine, Yale University School of Medicine, New Haven, Connecticut
    • Yale University School of Medicine, 333 Cedar St. (609 LCI), New Haven, CT 06510
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Abstract

RNA molecules immunoprecipitated with sera from patients who have rheumatic diseases can be readily detected in polyacrylamide gels by using ethidium bromide and silver stains. With these stains, we found that RNA patterns characteristic of a broad range of specific small nuclear ribonucleoproteins and small cytoplasmic ribonucleoproteins were recognizable. The stains correctly identified antibodies to ribonucleoproteins in 33 (92%) of 36 patient sera selected for study because of known antibody specificities. The silver stain method detected antibodies to ribonucleoproteins in 25 (76%) of 33 patients with classic systemic lupus erythematosus, a frequency that approximated the frequency observed in the Lerner-Steitz assay, which is based on autoradiography. This approach considerably simplifies the latter radioimmunoassay with minimal loss of precision and sensitivity.

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