Refolding Record:
| Protein | |
|---|---|
| Protein Name | Teratocarcinoma-derived growth factor 1 |
| Abbreviated Name | TDGF1 |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P13385 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Unknown |
| Construct | |
|---|---|
| Full Length | n |
| Domain | Epidermal growth factor-like domain aa.67-113 |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 47 |
| Molecular Weight | 5188.0 |
| Pi | 7.77 |
| Molecular Weight | 5188.0 |
| Disulphides | 3 |
| Full Sequence |
VPPMGIQHSKELNRTCCLNGGTCMLGSFCACPPSFYGRNCEHDVRKE
|
| Notes | CR47B construct |
| Expression | |
|---|---|
| Report | Lohmeyer M, Harrison PM, Kannan S, DeSantis M, OReilly NJ, Sternberg MJE, Salomon DS, Gullick WJ (1997) Biochemistry, 36, 3837-3845 |
| Project Aim | Structure-Function |
| Fusion | None |
| Protein Expression and Production | Protein synthesized by chemical means (not recombinant) and refolded. |
| Expression Host | None |
| Expression Strain | None |
| Expression Temp | 0.0 |
| Expression Time | n/a |
| Expression Vector | n/a |
| Expression Protocol | n/a |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | HPLC |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Dilution/Dialysis combination |
| Wash Buffer | n/a |
| Solubilization Buffer | 6.4M urea, 0.1M Na2SO3, 0.02M Na2S4O6 |
| Refolding Buffer | 1: 20mM Tris pH 7.1, 2: 13.3mM Tris, 0.03M sodium borate, 1mM EDTA pH 9.0 |
| Pre-Refolding Purification | HPLC |
| Tag Cleaved | no tag |
| Refolding pH | 9.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | 16-24h |
| Redox Agent | Cysteine |
| Redox Agent Concentration | 3mM,3mM,3mM |
| Refolding Protocol | Peptides were synthesised then purified by HPLC. The lyophilized peptide was dissolved in solubilization buffer, followed by sequential dialysis, firstly in Refolding buffer 1 with 1M urea, followed by Refolding buffer 1 alone. The peptide was then added to Refolding buffer 2 and incubated for 16-24h at room temperature. |
| Refolding Assay | Mass spectrometry |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |