Tandem affinity purification of functional TAP-tagged proteins from human cells.
Tandem affinity purification (TAP) is a generic two-step affinity purification protocol for isolation of TAP-tagged proteins together with associated proteins. We used bacterial artificial chromosome to heterologously express TAP-tagged murine Sgo1 protein in human HeLa cells. This allowed us to tes...
| Main Authors: | , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2007
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| _version_ | 1797068532885749760 |
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| author | Gregan, J Riedel, C Petronczki, M Cipak, L Rumpf, C Poser, I Buchholz, F Mechtler, K Nasmyth, K |
| author_facet | Gregan, J Riedel, C Petronczki, M Cipak, L Rumpf, C Poser, I Buchholz, F Mechtler, K Nasmyth, K |
| author_sort | Gregan, J |
| collection | OXFORD |
| description | Tandem affinity purification (TAP) is a generic two-step affinity purification protocol for isolation of TAP-tagged proteins together with associated proteins. We used bacterial artificial chromosome to heterologously express TAP-tagged murine Sgo1 protein in human HeLa cells. This allowed us to test the functionality of the Sgo1-TAP protein by RNA interference-mediated depletion of the endogenous human Sgo1. Here, we present an optimized protocol for purification of TAP-tagged Sgo1 protein as well as KIAA1387 from HeLa cells with detailed instructions. The purification protocol can be completed in 1 day and it should be applicable to other proteins. |
| first_indexed | 2024-03-06T22:11:57Z |
| format | Journal article |
| id | oxford-uuid:521d06a5-7244-446f-8b80-71ce4c93f974 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T22:11:57Z |
| publishDate | 2007 |
| record_format | dspace |
| spelling | oxford-uuid:521d06a5-7244-446f-8b80-71ce4c93f9742022-03-26T16:23:39ZTandem affinity purification of functional TAP-tagged proteins from human cells.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:521d06a5-7244-446f-8b80-71ce4c93f974EnglishSymplectic Elements at Oxford2007Gregan, JRiedel, CPetronczki, MCipak, LRumpf, CPoser, IBuchholz, FMechtler, KNasmyth, KTandem affinity purification (TAP) is a generic two-step affinity purification protocol for isolation of TAP-tagged proteins together with associated proteins. We used bacterial artificial chromosome to heterologously express TAP-tagged murine Sgo1 protein in human HeLa cells. This allowed us to test the functionality of the Sgo1-TAP protein by RNA interference-mediated depletion of the endogenous human Sgo1. Here, we present an optimized protocol for purification of TAP-tagged Sgo1 protein as well as KIAA1387 from HeLa cells with detailed instructions. The purification protocol can be completed in 1 day and it should be applicable to other proteins. |
| spellingShingle | Gregan, J Riedel, C Petronczki, M Cipak, L Rumpf, C Poser, I Buchholz, F Mechtler, K Nasmyth, K Tandem affinity purification of functional TAP-tagged proteins from human cells. |
| title | Tandem affinity purification of functional TAP-tagged proteins from human cells. |
| title_full | Tandem affinity purification of functional TAP-tagged proteins from human cells. |
| title_fullStr | Tandem affinity purification of functional TAP-tagged proteins from human cells. |
| title_full_unstemmed | Tandem affinity purification of functional TAP-tagged proteins from human cells. |
| title_short | Tandem affinity purification of functional TAP-tagged proteins from human cells. |
| title_sort | tandem affinity purification of functional tap tagged proteins from human cells |
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