Refolding Record:
| Protein | |
|---|---|
| Protein Name | Insulin-like growth factor binding protein 3 |
| Abbreviated Name | IGFBP-3 |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P17936 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Unknown |
| Construct | |
|---|---|
| Full Length | n |
| Domain | N terminal portion aa.1-147 |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 147 |
| Molecular Weight | 15093.1 |
| Pi | 5.90737 |
| Molecular Weight | 15093.1 |
| Disulphides | 0 |
| Full Sequence |
GAS SGGLGPVVRC EPCDARALAQ CAPPPAVCAE LVREPGCGCC LTCALSEGQP CGIYTERCGS GLRCQPSPDE ARPLQALLDG RGLCVNASAV SRLRAYLLPA PPAPGNASES EEDRSAGSVE
SPSVSSTHRV SDPKFHPLHS KIII
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Spencer EM and Chan K (1995) Progress in Growth Factor Research, 6, 209-214 |
| Project Aim | Structure-Function |
| Fusion | N-terminal Protein A |
| 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 | not stated |
| Expression Vector | pRIT2T |
| Expression Protocol | Protein was expressed in E.coli as a fusion protein with protein A. Inclusion bodies were harvested and solubilized. |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | Affinity chromotography |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | not stated |
| Refolding Buffer | DTT/cystamine redox system, pH 8.4 |
| Pre-Refolding Purification | Affinity chromotography |
| Tag Cleaved | yes |
| Refolding pH | 8.4 |
| Refolding Temperature | 25.0 |
| Protein Concentration | |
| Refolding Time | |
| Redox Agent | DTT/cystamine |
| Redox Agent Concentration | not stated |
| Refolding Protocol | After solubilization the fusion protein was partially purified by affinity chromatography using a gamma G sepharose column. Protein A was then cleaved from the protein by hydroxylamine and the unfolded protein was then reapplied to the gamma G Sepharose column to remove cleaved protein A and uncleaved fusion protein. The cleaved target protein was then refolded in a DTT/cystamine redox system at pH 8.4 under a N2 atmosphyere. |
| Refolding Assay | Western Blot |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | |
| Notes | |