Refolding Record:
| Protein | |
|---|---|
| Protein Name | Bovine serum albumin |
| Abbreviated Name | BSA |
| SCOP Family | Serum albumin-like |
| Structure Notes | |
| Organism | Bovine |
| UniProt Accession | P02769 |
| SCOP Unique ID | n/a |
| Structure Solved | |
| Class | Alpha |
| Molecularity | Monomer |
| Construct | |
|---|---|
| Full Length | n |
| Domain | n/a |
| Chimera | n/a |
| Variants | n/a |
| Chain Length | 583 |
| Molecular Weight | 66433.0 |
| Pi | 5.59869 |
| Molecular Weight | 66433.0 |
| Disulphides | >10 |
| Full Sequence |
DTHKSE IAHRFKDLGE EHFKGLVLIA FSQYLQQCPF DEHVKLVNEL TEFAKTCVAD ESHAGCEKSL HTLFGDELCK VASLRETYGD MADCCEKQEP ERNECFLSHK DDSPDLPKLK PDPNTLCDEF KADEKKFWGK YLYEIARRHP YFYAPELLYY ANKYNGVFQE CCQAEDKGAC LLPKIETMRE KVLASSARQR LRCASIQKFG ERALKAWSVA RLSQKFPKAE FVEVTKLVTD LTKVHKECCH GDLLECADDR ADLAKYICDN QDTISSKLKE CCDKPLLEKS HCIAEVEKDA IPENLPPLTA DFAEDKDVCK NYQEAKDAFL GSFLYEYSRR HPEYAVSVLL RLAKEYEATL EECCAKDDPH ACYSTVFDKL KHLVDEPQNL IKQNCDQFEK LGEYGFQNAL IVRYTRKVPQ VSTPTLVEVS RSLGKVGTRC CTKPESERMP CTEDYLSLIL NRLCVLHEKT PVSEKVTKCC TESLVNRRPC FSALTPDETY VPKAFDEKLF TFHADICTLP DTEKQIKKQT ALVELLKHKP KATEEQLKTV MENFVAFVDK CCAADDKEAC FAVEGPKLVV STQTALA
|
| Notes | n/a |
| Expression | |
|---|---|
| Report | Langenhof M, Leong SSJ, Pattenden LK, Middelberg APJ (2005) Journal of Chromatography A, 1069, 195-201 |
| Project Aim | Protein refolding |
| Fusion | None |
| Protein Expression and Production | Protein expressed and purified in native conformation prior to denaturation and refolding. |
| 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 | 50mM TrisHCl, 3mM EDTA, 8M urea, 0.1M DTT, pH 8.5 |
| Refolding Buffer | 50mM TrisHCl, 1mM EDTA, 1.1mM GSSG, 2.2mM GSH pH 8.5 |
| Pre-Refolding Purification | None |
| Tag Cleaved | no tag |
| Refolding pH | 8.5 |
| Refolding Temperature | 25.0 |
| Protein Concentration | 0.05-0.5mg/ml |
| Refolding Time | 96h |
| Redox Agent | GSH/GSSG |
| Redox Agent Concentration | 2.2/1.1mM,2.2/1.1mM |
| Refolding Protocol | The protein was denatured and reduced by incubating a 20-100mg/ml solution for at least 3h at room temperature in denaturation buffer. 2.5ml of the protein then underwent buffer exchange into denaturation buffer without DTT using a PD10 column. The protein was then diluted 100x into refolding buffer to a final concentration of 50micrg/ml-0.5mg/ml. The solution was incubated for 96h at room temperature. |
| Refolding Assay | Far-UV Circular Dichroism,HPLC |
| Refolding Chaperones | None |
| Refolding Additives | None |
| Additives Concentration | n/a |
| Refolding Yield | 82% |
| Purity | n/a |
| Notes | n/a |