Refolding Record:

Protein See all refolding records for this protein...
Protein Name	Suppressors of cytokine signalling
Abbreviated Name	SOCS
SCOP Family	SPRY domain
Structure Notes
Organism	Mouse
UniProt Accession	P14753
SCOP Unique ID	n/a
Structure Solved
Class	Beta
Molecularity	Unknown

Construct
Full Length	n
Domain	Construct lacks the first 22 residues of SOCS3 and is truncated before the SOCS box motif
Chimera	n/a
Variants	n/a
Chain Length	193
Molecular Weight	21422.9
Pi	6.82
Molecular Weight	21422.9
Disulphides	Unknown
Full Sequence	MASYHHHHHHHDYDIPTTENLYFQGAHDGSlktfsskseyqlvvnavrklqesgfywsavtggeanlllsaepagtflirdssdqrhfftlsvktqsgtknlriqceggsfslqsdprstqpvprfdcvlklvhhymppptpsfslpptepssevpeqppaqalpgstpkrayyiysggekiplvlsrplssn
Notes	n/a

Expression
Report	Babon JJ, Yao S, DeSouza DP, Harrison CF, Fabri LJ, Liepinsh E, Scrofani SD, Baca M, Norton RS. (2005) Febs Journal, 272, 6120-6130
Project Aim	Structural Studies
Fusion	N terminal hexa His tag + TEV cleavage sequence
Protein Expression and Production	Protein recombinantly expressed as and refolded from inclusion bodies.
Expression Host	Escherichia coli
Expression Strain	None
Expression Temp	30.0
Expression Time	3hrs
Expression Vector	Not stated
Expression Protocol	For unlabelled protein, expression was performed in baffled flasks, with cells grown to an attenuance (D) at 600 nm of 0.6 in superbroth containing 50 µg·mL1 kanamycin. Expression was induced with 1 mm IPTG. Cells were harvested, 3 h after induction, by centrifugation (6200 g, 4 °C, 30 min).
Method of Induction	IPTG
Cell Density at Induction	OD 600 = 0.6
Cell Disruption Method	High pressure homogenization
Lytic Agent	None
Pre-Refolding Purification	Metal affinity chromatography
Solubility	insoluble

Refolding
Refolding Method	Dialysis
Wash Buffer	Not stated
Solubilization Buffer	6M GuHCl
Refolding Buffer	25 mM sodium phosphate, 50 mM sodium chloride, 5 mM 2-mercaptoethanol, pH 6.7
Pre-Refolding Purification	Metal affinity chromatography
Tag Cleaved	no
Refolding pH	6.3
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 prepared via cell homogenization and centrifugation at 20 000 g, and solubilized using 6 m guanidine hydrochloride. The soluble inclusion body preparation was then purified using Ni-nitrilotriacetic acid resin (Qiagen, Valencia, CA, USA). Protein binding was performed at pH 8.0, washing at pH 6.3, and elution at pH 4.5. The eluted protein was quantified by absorbance at 280 nm, diluted to 0.1 mg·mL1, then refolded by extensive dialysis against 25 mM sodium phosphate, 50 mM sodium chloride, 5 mM 2-mercaptoethanol, pH 6.7.
Refolding Assay	Bioactivity
Refolding Chaperones	None
Refolding Additives	None
Additives Concentration	n/a
Refolding Yield	n/a
Purity	n/a
Notes	n/a

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