Refolding Record:
| Protein | |
|---|---|
| Protein Name | Major histocompatibility complex class I |
| Abbreviated Name | MHC-1 |
| SCOP Family | MHC antigen-recognition domain |
| Structure Notes | |
| Organism | Rat |
| UniProt Accession | O98177 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha+Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | n |
| Domain | n/a |
| Chimera | n/a |
| Variants | alpha chain |
| Chain Length | 84 |
| Molecular Weight | 9640.7 |
| Pi | 4.33 |
| Molecular Weight | 9640.7 |
| Disulphides | 0 |
| Full Sequence |
IKEEHTIIQAEFYLSPDQNGEFMFDFDGDEIFHVDIKKSETIWRLEEFAQFASFEAQGALANIAVDKANLDIMIKRSNNTPDAN
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Altman, J. D., Reay, P. A., Davis, M. M. (1993) Proc. Natl. Acad. Sci. USA, 90, 10330-10334 |
| Project Aim | Folding,Assembly |
| Fusion | None |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | Escherichia coli |
| Expression Strain | BL21(DE3)pLysS |
| Expression Temp | 37.0 |
| Expression Time | not stated |
| Expression Vector | pGeMEX-1 |
| Expression Protocol | no details given |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD n/a = n/a |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | insoluble |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | 6 M urea, 10mM acetic acid |
| Refolding Buffer | 50mM sodium phosphate, pH 7.5, 1mM EDTA, 3mM reduced glutathione, 0.3 mM oxidized glutathione, 25%(vol/vol) glycerol, 20 microM bioMCC. |
| Pre-Refolding Purification | None |
| Tag Cleaved | no |
| Refolding pH | 7.5 |
| Refolding Temperature | 4.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Inclusion bodies, were solubilized and run on a PD-10 desalting column. Refolding was initiated by the dilution in refolding buffer. |
| Refolding Assay | Complex Assembly |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |