Refolding Record:
| Protein | |
|---|---|
| Protein Name | Interferon gamma |
| Abbreviated Name | IFN gamma |
| SCOP Family | Interferons/interleukin-10 (IL-10) |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P01579 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha |
| Molecularity | Dimer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 140 |
| Molecular Weight | 16177.5 |
| Pi | 9.52198 |
| Molecular Weight | 16177.5 |
| Disulphides | 0 |
| Full Sequence |
QDPYVKEAENLKKYFNAGHSDVADNGTLFLGILKNWK
EESDRKIMQSQIVSFYFKLFKNFKDDQSIQKSVETIKEDMNVKFFNSNKKKRDDFEKLTN
YSVTDLNVQRKAIHELIQVMAELSPAAKTGKRKRSQMLFRG
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Geng Xindu, Bai Quan (2002) Science in China, 45, 655-669 |
| Project Aim | Folding |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | DH5¦Á |
| Expression Temp | 37.0 |
| Expression Time | 12 |
| Expression Vector | pBV 220 |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | Washing inclusion body |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Column refolding: hydrophobic interaction chromatography |
| Wash Buffer | 50mM PBS + 2 M urea |
| Solubilization Buffer | 7.0 M GuHCl +50mM PBS |
| Refolding Buffer | 50mM PBS |
| Pre-Refolding Purification | Washing inclusion body |
| Tag Cleaved | no tag |
| Refolding pH | 7.0 |
| Refolding Temperature | 25.0 |
| Protein Concentration | |
| Refolding Time | |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | The packed column, or USRPP was equilibrated with a new mobile phase, at least, for 15 min at each selected flow-rates before injecting a sample solution. The selection of flow rate for concentration gradient elution depended on the size of column or USRPP, and was detected at 280 nm. The eluted out fractions of the aim proteins were ollected for the measurements of the bioactivity and mass recoveries. Based on the various sample sizes injected required in various circumstances, the final concentrations of each proteins in the collected fractions were different. The start and final concentrations for protein refolding by HPHIC were listed in the notes of figure and table in the paper. |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | |
| Notes | |