Refolding Record:
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 129 |
| Molecular Weight | 14313.1 |
| Pi | 9.32374 |
| Molecular Weight | 14313.1 |
| Disulphides | Unknown |
| Full Sequence |
KVFGRCELAAAMKRHGLDNYRGYSLGNWVCAAKFESNFNTQATNRNTDGSTDYGILQINSRWWCNDGRTPGSRNLCNIPCSALLSSDITASVNCAKKIVSDGNGMNAWVAWRNRCKGTDVQAWIRGCRL
|
| Notes | Commercially available from Sigma Aldrich |
| Expression | |
|---|---|
| Report | Unpublished (0) J Virol Methods, 0, 0 |
| Project Aim | Folding |
| Fusion | None |
| Protein Expression and Production | Protein expressed and purified in native conformation prior to denaturation and refolding. |
| Expression Host | |
| Expression Strain | |
| Expression Temp | 0.0 |
| Expression Time | 0 |
| Expression Vector | |
| Expression Protocol | |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | None |
| Solubilization Buffer | 8 M urea in 50 mM phosphate, pH 8, 10 mM DTT |
| Refolding Buffer | Tris 50 mM, pH 8 with Schiff base reagents from Boston Protein Solutions |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | 0.91 mg/ml |
| Refolding Time | 20 hours |
| Redox Agent | GSH/GSSG |
| Redox Agent Concentration | 1 mM/2mM |
| Refolding Protocol | None. The Schiff reagents from Boston Protein Solutions are small molecule, non-detergent, water-soluble reagents. They function like regular salt, and dissociate from protein when being diluted and are easy to remove by dialysis and gel filtration. |
| Refolding Assay | Activity assay |
| Refolding Chaperones | None |
| Refolding Additives | Schiff base reagents from Boston Protein Solutions |
| Additives Concentration | 0.2%-2% |
| Refolding Yield | 87% |
| Purity | >90% |
| Notes | The Schiff reagents from Boston Protein Solutions can be used togetherwith other additives, like detergents, NDBS, arginine, proline etc. |