Refolding Record:
| Protein | |
|---|---|
| Protein Name | CD14 |
| Abbreviated Name | CD14 |
| SCOP Family | Unknown |
| Structure Notes | |
| Organism | Human |
| UniProt Accession | P08571 |
| 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 | 381 |
| Molecular Weight | 40076.2 |
| Pi | 5.84457 |
| Molecular Weight | 40076.2 |
| Disulphides | Unknown |
| Full Sequence |
MERASCLLLLLLPLVHVSATTPEPCELDDEDFRCVCNFSEPQPDWSEAFQCVSAVEVEIH
AGGLNLEPFLKRVDADADPRQYADTVKALRVRRLTVGAAQVPAQLLVGALRVLAYSRLKE
LTLEDLKITGTMPPLPLEATGLALSSLRLRNVSWATGRSWLAELQQWLKPGLKVLSIAQA
HSPAFSCEQVRAFPALTSLDLSDNPGLGERGLMAALCPHKFPAIQNLALRNTGMETPTGV
CAALAAAGVQPHSLDLSHNSLRATVNPSAPRCMWSSALNSLNLSFAGLEQVPKGLPAKLR
VLDLSCNRLNRAPQPDELPEVDNLTLDGNPFLVPGTALPHEGSMNSGVVPACARSTLSVG
VSGTLVLLQGARGFA
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Majerle A, Kidric J, Jerala R. (2000) Pflugers Arch, 439, 109-110 |
| 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 | 37.0 |
| Expression Time | 3h |
| Expression Vector | pET3a |
| 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 | 10mM Tris-HCl, 0.5mM EDTA, 0.1% deoxycholate, pH 8.0 |
| Solubilization Buffer | 6M guanidinium chloride, 20mM Tris-HCl, 20mM DTT pH 8.0 |
| Refolding Buffer | 20mM Tris-HCl, 5mM cysteine, 3mM EDTA |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.0 |
| Refolding Temperature | 4.0 |
| Protein Concentration | 0.02mg/ml |
| Refolding Time | 18h |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | Inclusion bodies were washed twice in 10mM Tris-HCl, 0.5mM EDTA, 0.1% deoxycholate, pH 8.0 followed by one wash with the same buffer also containing 2M urea. The protein was solubilized in 6M guanidinium chloride, 20mM Tris-HCl, 20mM DTT pH 8.0 and refolded by rapid dilution. |
| Refolding Assay | Immunoassay |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | NULL |
| Refolding Yield | |
| Purity | |
| Notes | |