Refolding Record:
| Protein | |
|---|---|
| Protein Name | Growth Hormone |
| Abbreviated Name | GH |
| SCOP Family | Long-Chain Cytokines |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P01241 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 191 |
| Molecular Weight | 22129.0 |
| Pi | 5.268 |
| Molecular Weight | 22129.0 |
| Disulphides | 2 |
| Full Sequence |
FPTIPLSRLFDNAMLRAHRLHQLAFDTYQEFEEAYIPKEQKYSFLQNPQTSLCFSESIPTPSNREETQQKSNLELLRISLLLIQSWLEPVQFLRSVFANSLVYGASDSNVYDLLKDLEEGIQTLMGRLEDGSPRTGQIFKQTYSKFDTNSHNDDALLKNYGLLYCFRKDMDKVETFLRIVQCRSVEGSCGF
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | St John, R.J., Carpenter, J.F., Claude, B., Randolph, T.W. (2001) J Biol Chem, 276, 46856-46863 |
| Project Aim | Folding |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | None |
| Expression Temp | 37.0 |
| Expression Time | unknown |
| Expression Vector | unknown |
| Expression Protocol | Not stated |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | High Pressure |
| Wash Buffer | unknown |
| Solubilization Buffer | unknown |
| Refolding Buffer | 10 mM sodium citrate, 1 mM EDTA |
| Pre-Refolding Purification | None |
| Tag Cleaved | yes |
| Refolding pH | 6.0 |
| Refolding Temperature | 65.0 |
| Protein Concentration | 0.65 mg/ml |
| Refolding Time | 24 hrs |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Agitation induced aggregates of human growth hormone (Genenech) were diluted in refolding buffer and placed in a heat sealed plastic syringe. The samples were placed under pressure at 2000 bar then heated to 65 deg C and held for 24 hours, cooled then depressurized slowly. |
| Refolding Assay | Ultraviolet (UV) Absorbance |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 90% |
| Purity | |
| Notes | |