Refolding Record:
| Protein | |
|---|---|
| Protein Name | Heparin-binding neurite-promoting factor |
| Abbreviated Name | HBNF |
| SCOP Family | Midkine, a heparin-binding growth factor, C-terminal domain |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P21246 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Small Proteins |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 170 |
| Molecular Weight | 18942.0 |
| Pi | 9.66222 |
| Molecular Weight | 18942.0 |
| Disulphides | 2 |
| Full Sequence |
MQAQQYQQQRRKFAAAFLAFIFILAAVDTAEAGKKEKPEKKVKKSDCGEWQWSVCVPTSG
DCGLGTREGTRTGAECKQTMKTQRCKIPCNWKKQFGAECKYQFQAWGECDLNTALKTRTG
SLKRALHNAECQKTVTISKPCGKLTKPKPQAESKKKKKEGKKQEKMLD
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Seddon AP, Hulmes JD, Decker MM, Kovesdi I, Fairhurst JL, Backer J, Dougher-Vermazen M, Bohlen P. Seddon AP, Hulmes JD, Decker MM, Kovesdi I, Fairhurst JL, Backer J, Dougher-Vermazen M, Bohlen P. (1994) Protein Expression and Purification, 5, 14-21 |
| Project Aim | Undefined |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21 |
| Expression Temp | 37.0 |
| Expression Time | 2h |
| Expression Vector | pET3a |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | French Press |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | unknown |
| Solubilization Buffer | 50 mM Tris-HCl, 10mM DTT, 0.1mM EDTA, 1M NaCl, 8M urea pH 7.5 |
| Refolding Buffer | 25mM Hepes, 1M NaCl |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 7.5 |
| Refolding Temperature | 4.0 |
| Protein Concentration | 1mg/ml |
| Refolding Time | 16h |
| Redox Agent | DTT |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The solubilized protein from 1L culture was dialysed against 4L of buffer for 8h, followed by a second identical dialysis. |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | 91% |
| Purity | 85% |
| Notes | |