HOXB13 Interactome Profiling by Mass Spectrometry
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Corresponding Organization :
Other organizations : Northwestern University, Dana-Farber Cancer Institute, University of Washington
Variable analysis
- Stable expression of HOXB13 WT-SFB or G84E-SFB in LNCaP cells
- Proteins identified and sequenced by mass spectrometry analysis
- Cell lysis in NETN buffer containing protease inhibitors
- Centrifugation at 21,100×g for 30 min
- Incubation with streptavidin-conjugated beads for 4 h at 4 °C
- Washing of beads with NETN buffer
- Elution of bounded proteins with NETN buffer containing biotin
- Incubation with S-protein beads overnight at 4 °C
- Elution of proteins with SDS sample buffer
- SDS-PAGE separation of proteins
- Protein identification and peptide sequencing using SEQUEST
- Positive control: Not specified
- Negative control: Not specified
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