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- Publisher Website: 10.1038/nprot.2007.492
- Scopus: eid_2-s2.0-38149003098
- PMID: 18193021
- WOS: WOS:000253140400006
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Article: Southwestern blotting in investigating transcriptional regulation
Title | Southwestern blotting in investigating transcriptional regulation |
---|---|
Authors | |
Issue Date | 2008 |
Publisher | Nature Publishing Group. The Journal's web site is located at http://www.natureprotocols.com/about.php |
Citation | Nature Protocols, 2008, v. 3 n. 1, p. 51-58 How to Cite? |
Abstract | Southwestern blotting is used to investigate DNA-protein interactions. The advantage of this technique over other related methods such as electrophoretic mobility shift assay (EMSA) and DNA footprinting is that it provides information regarding the molecular weight of unknown protein factor. This method combines the features of Southern and Western blotting techniques; a denaturing SDS-PAGE is first employed to separate proteins electrophoretically based on size, and after transferring the proteins to a membrane support, the membrane-bound proteins are renatured and incubated with a 32P-labeled double-stranded oligonucleotide probe of specific DNA sequence. The interaction of the probe with the protein(s) is later visualized by autoradiography. This technique could be combined with database searching (TransFac, http://www.gene-regulation.com/pub/ databases.html#transfac), prediction of potential protein factors binding onto a target motif (e.g., Patch search), in vitro supershift EMSA and in vivo chromatin immunoprecipitation (ChIP) assays for effective identification of protein factors. The whole Southwestern blotting procedure takes ∼4 d to complete. In this article, a commonly used protocol and expected results are described and discussed. |
Persistent Identifier | http://hdl.handle.net/10722/89291 |
ISSN | 2023 Impact Factor: 13.1 2023 SCImago Journal Rankings: 7.419 |
ISI Accession Number ID | |
References |
DC Field | Value | Language |
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dc.contributor.author | Siu, FKY | en_HK |
dc.contributor.author | Lee, LTO | en_HK |
dc.contributor.author | Chow, BKC | en_HK |
dc.date.accessioned | 2010-09-06T09:55:00Z | - |
dc.date.available | 2010-09-06T09:55:00Z | - |
dc.date.issued | 2008 | en_HK |
dc.identifier.citation | Nature Protocols, 2008, v. 3 n. 1, p. 51-58 | en_HK |
dc.identifier.issn | 1754-2189 | en_HK |
dc.identifier.uri | http://hdl.handle.net/10722/89291 | - |
dc.description.abstract | Southwestern blotting is used to investigate DNA-protein interactions. The advantage of this technique over other related methods such as electrophoretic mobility shift assay (EMSA) and DNA footprinting is that it provides information regarding the molecular weight of unknown protein factor. This method combines the features of Southern and Western blotting techniques; a denaturing SDS-PAGE is first employed to separate proteins electrophoretically based on size, and after transferring the proteins to a membrane support, the membrane-bound proteins are renatured and incubated with a 32P-labeled double-stranded oligonucleotide probe of specific DNA sequence. The interaction of the probe with the protein(s) is later visualized by autoradiography. This technique could be combined with database searching (TransFac, http://www.gene-regulation.com/pub/ databases.html#transfac), prediction of potential protein factors binding onto a target motif (e.g., Patch search), in vitro supershift EMSA and in vivo chromatin immunoprecipitation (ChIP) assays for effective identification of protein factors. The whole Southwestern blotting procedure takes ∼4 d to complete. In this article, a commonly used protocol and expected results are described and discussed. | en_HK |
dc.language | eng | en_HK |
dc.publisher | Nature Publishing Group. The Journal's web site is located at http://www.natureprotocols.com/about.php | en_HK |
dc.relation.ispartof | Nature Protocols | en_HK |
dc.title | Southwestern blotting in investigating transcriptional regulation | en_HK |
dc.type | Article | en_HK |
dc.identifier.openurl | http://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1750-2799&volume=3&spage=51&epage=58&date=2008&atitle=Southwestern+blotting+in+investigating+transcriptional+regulation | en_HK |
dc.identifier.email | Lee, LTO: ltolee2@hkucc.hku.hk | en_HK |
dc.identifier.email | Chow, BKC: bkcc@hku.hk | en_HK |
dc.identifier.authority | Lee, LTO=rp00727 | en_HK |
dc.identifier.authority | Chow, BKC=rp00681 | en_HK |
dc.description.nature | link_to_subscribed_fulltext | - |
dc.identifier.doi | 10.1038/nprot.2007.492 | en_HK |
dc.identifier.pmid | 18193021 | - |
dc.identifier.scopus | eid_2-s2.0-38149003098 | en_HK |
dc.identifier.hkuros | 140748 | en_HK |
dc.relation.references | http://www.scopus.com/mlt/select.url?eid=2-s2.0-38149003098&selection=ref&src=s&origin=recordpage | en_HK |
dc.identifier.volume | 3 | en_HK |
dc.identifier.issue | 1 | en_HK |
dc.identifier.spage | 51 | en_HK |
dc.identifier.epage | 58 | en_HK |
dc.identifier.eissn | 1750-2799 | - |
dc.identifier.isi | WOS:000253140400006 | - |
dc.identifier.scopusauthorid | Siu, FKY=6701518484 | en_HK |
dc.identifier.scopusauthorid | Lee, LTO=8367269000 | en_HK |
dc.identifier.scopusauthorid | Chow, BKC=7102826193 | en_HK |
dc.identifier.citeulike | 2761685 | - |
dc.identifier.issnl | 1750-2799 | - |