| Species: | Human |
Expression Host: | Human 293HEK cells |
| Expression cDNA Clone and Sequence |
Recombinant protein was produced with TrueORF clone, RC208093. Click on the TrueORF clone link to view cDNA and protein sequences. |
| Tag: | C-terminal MYC/DDK |
Predicted MW: | 53.7 kDa |
| Purity: | > 80% as determined by SDS-PAGE and Coomassie blue staining |
| Mass Spec Validation: |
This protein has been positively validated by MS/MS by Dr. Robert Moritz at The Institute of Systems Biology. |
| Concentration: | >50 ug/mL as determined by microplate BCA method |
| Buffer and Storage: | 25 mM Tris.HCl, pH 7.3, 100 mM glycine, 10% glycerol. Store at -80C. Avoid repeated freeze-thaw cycles. Stable for at least 3 months from receipt of products under proper storage and handling conditions. |
| Preparation: |
Recombinant protein was captured through anti-DDK affinity column followed by conventional chromatography steps. |
| BioActivity: |
Varying amounts of human ERG expressed in HEK293 cells was incubated for one hour with wild-type or mutant biotinylated oligonucleotide (1 pmole/ul) in the presence of 25 ug/ml poly dI:dC. The reaction mixture was subsequently transferred to a microplate containing 2500 Luminex beads coupled with anti-ERG monoclonal antibody 2G8. The ERG-oligo complexes were captured onto the antibody-coated beads for two hours at room temperature with shaking. The beads were then washed, and the biotin was detected with streptavidin-phycoerythrin for 30 minutes. The beads were washed again and the fluorescent intensity was read in the Luminex instrument. The wild-type oligonucleotide carried ACCGGAAGT consensus binding sequence while the mutant oligonucleotide was identical except for a 2-base mutation in the consensus binding region, ACCCCAAGT |
| Protein Families: |
Transcription FactorsDruggable Genome |
| Protein Pathways: |
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