| Species: | Human |
Expression Host: | Human 293HEK cells |
| Expression cDNA Clone and Sequence |
Recombinant protein was produced with TrueORF clone, RC217863. Click on the TrueORF clone link to view cDNA and protein sequences. |
| Tag: | C-terminal MYC/DDK |
Predicted MW: | 52.6 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: |
HNF4A Activity Verified in a DNA-binding Assay: HNF4A (TP317863, transcript variant 2) activity was measured in a colorimetric DNA-binding assay. Purified HNF4A protein containing a C-terminal MYC/DDK tag was incubated with biotinylated double-stranded oligonucleotide containing the HNF4A consensus DNA-binding sequence (see below). Following incubation, the reaction was transferred to a streptavidin-coated microplate to allow capture of the DNA-protein complex. After washing, the captured protein was detected with an anti-DDK peroxidase conjugate and colorimetric signal detection with TMB. Specificity of the protein-DNA interaction was confirmed by carrying out the binding in the presence of an unlabeled competitor oligonucleotide and by comparison to binding to an oligonucleotide containing a mutation in the consensus binding sequence. |
| Protein Families: |
ES Cell Differentiation/IPSNuclear Hormone ReceptorTranscription FactorsDruggable Genome |
| Protein Pathways: |
Maturity onset diabetes of the young |
