Send to:

Choose Destination
See comment in PubMed Commons below
Biochem Biophys Res Commun. 1993 Feb 26;191(1):18-25.

RNA-binding patterns in total human tissue proteins: analysis by northwestern blotting.

Author information

  • 1Department of Neurology, Columbia University College of Physicians and Surgeons, New York, NY 10032.


We have developed a reproducible Northwestern (NW) blotting method to identify general patterns of RNA-binding proteins in total human tissue homogenates and have identified some of the factors contributing to this reproducibility. Unfractionated homogenates of human brain tissue were separated by SDS-PAGE, electrotransferred wet to nitrocellulose, and probed with in-vitro-transcribed labeled RNAs. Approximately ten size classes of RNA-binding proteins were observed consistently and reproducibly. Although sequence-independent electrostatic RNA-protein interactions likely contributed to most of the binding, binding to some proteins was shown to be more dependent on protein conformation: binding was not blocked by preincubation with single- or double-stranded DNA, nor with poly(A) RNA, but preincubation with tRNA revealed a distinct subset of RNA-binding proteins. In addition, preincubation with RNA, but not DNA, revealed a previously undetected RNA-binding protein of approximately 90 kDa. The NW blotting method described here can be used to reveal tissue-specific differences in RNA-binding patterns.

[PubMed - indexed for MEDLINE]
PubMed Commons home

PubMed Commons

How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science
    Loading ...
    Write to the Help Desk