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StrepTactin Sepharose High Performance/StrepTrap HP

• Highly pure Strep(II)-tagged¹ recombinant proteins eluted in concentrated form and small volumes.
• StrepTrap HP 1 ml and 5 ml columns prepacked with StrepTactin™ Sepharose™ High Performance give convenient, time-saving operation and reproducible results.
• Physiological conditions and mild elution preserves the activity of the target protein.
• Compatible with a wide range of reducing agents, detergents, denaturants, and other additives.
• Easy scale-up and easily regenerated with 0.5 M NaOH.
• Convenient use with a peristaltic pump, or chromatography system such as ÄKTAdesign™.

StrepTactin Sepharose High Performance/StrepTrap HP

Technical Information

StrepTactin Sepharose High Performance and StrepTrap HP 1 ml and 5 ml columns for purification of Strep(II)-tagged recombinant proteins to high purities in concentrated forms and small volumes.

StrepTrap™ HP is supplied ready to use for convenient purification of Strep(II)-tagged recombinant proteins by affinity chromatography. These columns are prepacked with StrepTactin Sepharose High Performance. This robust and stable Sepharose based medium results in high resolution, sharp concentrated peaks, and reproducible purifications. The binding capacity is also high.

Purification is performed under physiological conditions and the mild elution preserves the activity of the target protein. Due to the high specificity of the binding, very high purity of the eluted protein is achieved in just one step. Regeneration of StrepTactin Sepharose High Performance and StrepTrap HP is fast and easily performed with 0.5 M NaOH.

Six repetitive GAPDH-Strep(II) purification runs on the same StrepTrap HP 1 ml column including regeneration with 0.5 M NaOH between each run. For SDS-PAGE analysis and recovery data, see next page.

SDS-PAGE (reducing conditions) of the six repetitive purifications runs in the regeneration study shows excellent reproducibility.

Recovery in the eluted fractions was retained over six purification runs.

COMPATABILITY GUIDE - Compatibility of StrepTactin Sepharose High Performance with different additives*
Reagent	Concentration
Reducing agents
DTT	50 mM
b-mercaptoethanol	50 mM
Nonionic detergents
C8E4, Octyltetraoxyethylene	max. 0.88%
C10E5, Decylpentaoxyethylene	0.12%
C10E6	0.03%
C12E8	0.01%
C12E9, Dodecyl nonaoxyethylene (Thesit)	0.02%
Decyl-b-D-maltoside	0.35%
N-dodecyl-b-D-maltoside	0.01%
N-nonyl-b-D-glucopyranoside	0.20%
N-octyl-b-D-glucopyranoside	2.34%
Triton X-100	2%
Tween 20	2%
Ionic detergents
N-lauryl-sarcosine	2%
8-HESO; N-octyl-2-hydroxy-ethylsulfoxide	1.32%
SDS, Sodium-N-dodecyl sulfate	0.10%
Zwitterionic detergents
CHAPS	0.10%
DDAO, N-decyl-N,N-dimethylamine-N-oxide	0.03%
LDAO, N-dodecyl-N,N-dimethylamine-N-oxide	0.13%
Others
Ammonium sulfate, (NH4)2SO4	2 M
CaCl2	max. 1 M
EDTA	50 mM
Guanidine	max. 1 M
Glycerol	max. 25%†
Imidazole	500 mM‡
MgCl2	1 M
Urea	max. 1 M
NaCl	5 M
These reagents have been successfully tested for the purification of, for example, GAPDH-Strep(II) with concentrations up to those listed. For reagents not marked with 'max', higher concentrations may be possible. Binding depends on the sterical accessibility of the Strep(II) tag and therefore varies from protein to protein. * Data kindly provided by IBA GmbH, Germany, the manufacturer and intellectual property owner of the StrepTactin ligand. † Yield may decrease. ‡ 500 mM imidazole in sample tested by GE Healthcare.

TECHNICAL SPECIFICATIONS
Matrix	Rigid, highly cross-linked 6% agarose
Average particle size	34 μm
Ligand	StrepTactin
Dynamic binding capacity1	Approx. 6 mg Strep(II)-tagged protein/ml medium
Compatibility	Stable in all commonly used buffers, 0.5 M NaOH (regeneration and cleaning), reducing agents, denaturants, and detergents, see table on next page for more information
pH stability	> 7
Storage	4°C to 8°C in 20% ethanol
StrepTrap HP
Column volume	1 ml and 5 ml
Column dimensions	0.7 × 2.5 cm (1ml); 1.6 × 2.5 cm (5 ml)
Recommended flow rate2	1 ml/min (1 ml); 5 ml/min (5 ml)
Max. flow rate2	4 ml/min (1 ml); 20 ml/min (5 ml)
Max. pressure2	0.3 MPa, 3 bar
Storage	20% ethanol
Storage temperature	4°C to 8°C
1 Binding capacity is protein dependent. Dynamic binding capacity was tested with GAPDH-Strep(II), Mr 37 400) and MBP-bGal (MBP2*-beta-galactosidase, Mr approx. 158 000, multimer in solution) 2 H20 at room temperature

详情请查询以下网址：http://www.gelifesciences.com/aptrix/upp01077.nsf/Content/Products?OpenDocument&moduleid=167321