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 | 168 |
| Molecular Weight | 19348.3 |
| Pi | 9.49784 |
| Molecular Weight | 19348.3 |
| Disulphides | 0 |
| Full Sequence |
MKYTSYILAFQLCIVLGSLGCYCQDPYVKEAENLKKYFNAGHSDVADNGTLFLGILKNWK
EESDRKIMQSQIVSFYFKLFKNFKDDQSIQKSVETIKEDMNVKFFNSNKKKRDDFEKLTN
YSVTDLNVQRKAIHELIQVMAELSPAAKTGKRKRSQMLFRGRRASQ
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Arora D, Khanna N. (1996) J Biotechnology, 52, 127-133 |
| 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 | 42.0 |
| Expression Time | 4h |
| Expression Vector | unknown |
| Expression Protocol | unknown |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | 100mM Tris-HCl, 10mM EDTA, 100mM NaCl, 4M urea, 0.5mM PMSF, pH 8.0 |
| Solubilization Buffer | 6M guanidinium chloride, 0.1M Tris-HCl, 0.2mM EDTA, pH 8.0 |
| Refolding Buffer | 0.1M Tris-HCl, 0.2mM EDTA, 0.5M L-arginine |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.0 |
| Refolding Temperature | 10.0 |
| Protein Concentration | |
| Refolding Time | 36-48h |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Three injections of denatured inclusion bodies were made into the refolding buffer 2h apart. The solution was stirred briefly, and incubated at 10C without further stirring either until a another addition of denatured inclusion bodies or the end of incubation. |
| Refolding Assay | Bioactivity |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | |
| Notes | |