Refolding Record:
| Protein | |
|---|---|
| Protein Name | Ciliary neurotrophic factor receptor alpha |
| Abbreviated Name | CNTFRalpha |
| SCOP Family | Fibronectin type III |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P26992 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 353 |
| Molecular Weight | 38897.1 |
| Pi | 6.26337 |
| Molecular Weight | 38897.1 |
| Disulphides | 1 |
| Full Sequence |
MYAQRHSPQEA PHVQYERLGS DVTLPCGTAN WDAAVTWRVN GTDLAPDLLN GSQLVLHGLE LGHSGLYACF HRDSWHLRHQ VLLHVGLPPR EPVLSCRSNT YPKGFYCSWH LPTPTYIPNT FNVTVLHGSK IMVCEKDPAL KNRCHIRYMH LFSTIKYKVS ISVSNALGHN ATAITFDEFT IVKPDPPENV VARPVPSNPR RLEVTWQTPS TWPDPESFPL KFFLRYRPLI LDQWQHVELS DGTAHTITDA YAGKEYIIQV AAKDNEIGTW SDWSVAAHAT
PWTEEPRHLT TEAQAAETTT STTSSLAPPP TTKICDPGEL GSGGGPSAPF LVSVPITLAL AAAAATASSL LI
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Panayotatos N, Everdeen D, Liten A, Somogyi R, Acheson A (1994) Biochemistry, 33, 5813-5818 |
| Project Aim | Functional Studies |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | K12 |
| Expression Temp | 37.0 |
| Expression Time | not stated |
| Expression Vector | not stated |
| Expression Protocol | no detail provided |
| 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 | Dialysis |
| Wash Buffer | n/a |
| Solubilization Buffer | 8M GdnHCl |
| Refolding Buffer | 10mM TrisHCl, 5mM EDTA pH 8.5, 0.6mg/ml GSH, 0.12mg/ml GSSG |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.5 |
| Refolding Temperature | 4.0 |
| Protein Concentration | |
| Refolding Time | 24h |
| Redox Agent | GSH/GSSG |
| Redox Agent Concentration | 0.6mM/0.12mM |
| Refolding Protocol | The protein was extracted from inclusion bodies in solubilization buffer, then dialyzed against refolding buffer for 24h at 4degC. The refolded protein was filtered then purified further using a Sephacryl S-100 column, then a Porous II/Q/P column. |
| Refolding Assay | Ligand Binding |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | |
| Refolding Yield | |
| Purity | 95% |
| Notes | |