Refolding Record:
| Protein | |
|---|---|
| Protein Name | non-POU domain-containing octamer binding protein |
| Abbreviated Name | NonO |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Rat |
| UniProt Accession | Q5FVM4 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Unknown |
| Molecularity | Unknown |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 510 |
| Molecular Weight | 58860.6 |
| Pi | 8.33 |
| Molecular Weight | 58860.6 |
| Disulphides | Unknown |
| Full Sequence |
MTMITPSLKDHLIHNVHKEEHAHAHNKIDDDDKVNQSNKTFNLEKQNHTPRKHHQHHHQQHHQQQQQQQQQQQQQPPPPIPANGQQASSQNEGLTIDLKNFRKPGEKTFTQRSRLFVGNLPPDITEEEMRKLFEKYGKAGEVFIHKDKGFGFIRLETRTLAEIAKVELDNMPLRGKQLRVRFACHSASLTVRNLPQYVSNELLEEAFSVFGQVERAVVIVDDRGRPSGKGIVEFSGKPAARKALDRCSEGSFLLTTFPRPVTVEPMDQLDDEEGLPEKLVIKNQQFHKEREQPPRFAQPGSFEYEYAMRWKALIEMEKQQQDQVDRNIKEAREKLEMEMEAARHEHQVMLMRQDLMRRQEELRRMEELHNQEVQKRKQLELRQEEERRRREEEMRRQQEEMMRRQQEGFKGTFPDAREQEIRMGQMAMGGAMGINNRGAMPPAPVPPGTPAPPGPAAMMPDGTLGLTPPTTERFGQAATMEGIGAIGGTPPAFNRPAPGAEFAPNKRRRY
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Unpublished (0) J Virol Methods, 0, 0 |
| Project Aim | Protein-Protein Interaction Identification |
| Fusion | N-terminal hexahistidine tag |
| Protein Expression and Production | Protein recombinantly expressed as and refolded from inclusion bodies. |
| Expression Host | E coli |
| Expression Strain | BL21 |
| Expression Temp | 37.0 |
| Expression Time | 2h - 12h |
| Expression Vector | pHAT |
| Expression Protocol | Ni-resin affinity |
| Method of Induction | IPTG |
| Cell Density at Induction | OD 0.7 = 600 |
| Cell Disruption Method | Sonication |
| Lytic Agent | None |
| Pre-Refolding Purification | Affinity chromotography |
| Solubility | soluble |
| Refolding | |
|---|---|
| Refolding Method | Dialysis |
| Wash Buffer | 8M Urea, 100mM potassium phosphate buffer,pH7.2 |
| Solubilization Buffer | 8M Urea, 100mM potassium phosphate buffer,pH7.2 |
| Refolding Buffer | 150mM NaCl, 50mM potassium phosphate buffer,pH8.0, 10%glycerol, 0.05% Tween 20, 2mM 2-mercaptpethanol |
| Pre-Refolding Purification | Affinity chromotography |
| Tag Cleaved | no |
| Refolding pH | 8.0 |
| Refolding Temperature | 4.0 |
| Protein Concentration | 1 mg/ml |
| Refolding Time | 4h |
| Redox Agent | Beta-mercaptoethanol |
| Redox Agent Concentration | n/a,n/a |
| Refolding Protocol | dialysis |
| Refolding Assay | Electrophoretic mobility-shift assays |
| Refolding Chaperones | None |
| Refolding Additives | tween 20 |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |