Refolding Record:
| Protein | |
|---|---|
| Protein Name | Carbonic Anhydrase B |
| Abbreviated Name | CAB |
| SCOP Family | Carbonic anhydrase |
| Structure Notes | |
| Organism | Bovine |
| UniProt Accession | P00921 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Beta |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | y |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 303 |
| Molecular Weight | 33430.7 |
| Pi | 8.49 |
| Molecular Weight | 33430.7 |
| Disulphides | Unknown |
| Full Sequence |
METQVSSRSTSEPVGCQHPKGAENVALISLAKSCDRPSSFRKPRELRQYTNMEASPEHWGKLYPIANGNNQSPIDIKTSETKRDPSLKPLSVSYNPATAKEIVNVGHSFHVNFEDSDNRSVLKGGPLSESYRLRQFHFHWGITDDCGSEHLVDGAKFSAELHLVHWNSAKYTSFADAASQADGLALIGVLVKVGQANPNLQKVLDALKAVKNKNKKAPFTNFDPSVLLPPSLDYWAYSGSLTHPPLHESVTWIIFKETISASSEQLAQFRCLLANAEGDGELCIKQNHRPPQPLKGRTVKASF
|
| Notes | CAB purchased from Sigma |
| Expression | |
|---|---|
| Report | Chen, Y., Huang, L. W., Chui, H. C., Lin, S. C. (2003) Enzyme Microb. Technol., 32, 120-130 |
| Project Aim | Folding |
| Fusion | None |
| Protein Expression and Production | Protein synthesized by chemical means (not recombinant) and refolded. |
| Expression Host | None |
| Expression Strain | None |
| Expression Temp | 0.0 |
| Expression Time | 0 |
| Expression Vector | |
| Expression Protocol | |
| Method of Induction | Not Stated |
| Cell Density at Induction | OD = |
| Cell Disruption Method | None |
| Lytic Agent | None |
| Pre-Refolding Purification | None |
| Solubility | |
| Refolding | |
|---|---|
| Refolding Method | Dilution |
| Wash Buffer | n/a |
| Solubilization Buffer | 20mM Tris-sulfate pH 7.5, 5mM EDTA, 5M GdnHCl |
| Refolding Buffer | 50mM Tris-sulfate pH 7.5, 5mM EDTA |
| Pre-Refolding Purification | None |
| Tag Cleaved | no |
| Refolding pH | 7.5 |
| Refolding Temperature | 25.0 |
| Protein Concentration | n/a |
| Refolding Time | n/a |
| Redox Agent | None |
| Redox Agent Concentration | n/a |
| Refolding Protocol | CAb was denatured using solubilisation buffer and incubated at room temp for 12h. It was then refolded using refolding buffer with PEG 4000, and PNIPAAm. |
| Refolding Assay | Fluorescence |
| Refolding Chaperones | None |
| Refolding Additives | Polyethylene glycol (PEG),PNIPAAm |
| Additives Concentration | n/a |
| Refolding Yield | n/a |
| Purity | n/a |
| Notes | n/a |