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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Rio, Donald C. |
| Abstract | It is of interest to be able to define sets of cellular RNAs associated with specific RNA-binding proteins. This 'guilt by association' can lead to new insights into how RNA-binding proteins modulate posttranscriptional gene expression of specific target RNAs. To identify these RNAs, antibodies against RNA-binding proteins can be used to immunopurify endogenous RNA-protein complexes from cells, and then the associated RNAs can be characterized. The method described here was developed to identify binding regions on nuclear transcripts for Drosophila heterogeneous nuclear ribonucleoproteins (hnRNPs). An antibody is added to an RNP extract and incubated to allow antigen-antibody complexes to form. The antibody-antigen complexes are then retrieved by binding of the antibody constant region to staphylococcal Protein A immobilized on Sepharose beads. The bead-immobilized complexes are then washed and RNA is prepared. The RNA is used to generate random-primed cDNA, cRNA, and biotinlyated cDNA probes for use on Affymetrix whole-genome Drosophila tiling arrays. |
| File Format | HTM / HTML |
| ISSN | 19403402 |
| Issue Number | 3 |
| Volume Number | 2014 |
| e-ISSN | 15596095 |
| Journal | Cold Spring Harbor Protocols |
| Language | English |
| Publisher | Cold Spring Harbor Laboratory Press |
| Publisher Date | 2014-03-01 |
| Publisher Place | United States |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Clinical Laboratory Techniques Cell Nucleus Metabolism Cytoplasm Drosophila Oligonucleotide Array Sequence Analysis Ribonucleoproteins Isolation & Purification Animals Antigen-antibody Complex Journal Article |
| Content Type | Text |
| Resource Type | Article |
| Subject | Biochemistry, Genetics and Molecular Biology |
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