IBA : Protein Tools

for purification of Strep-tag^® II proteins by gravity-flow column chromatography

Strep-Tactin Sepharose provides high binding capacity and minimal non-specific binding (see figure below). The material can be used for gravity flow purification. Since the Strep-Tactin resins are optimized for column affinity chromatography as opposed to batch purification, we do recommend our pre-packed Strep-Tactin Sepharose columns, which are available in different formats (from 0.2 ml mini columns up to 10 ml columns). Further specifications on the various column formats can be found here. For packing individual columns bulk material is available.

In one step: over 99% purity and high purification factors

A 36 kDa enzyme and a mutant, both with C-terminal Strep-tag II, were expressed in the cytoplasm of E. coli. The crude lysate was chromatographically separated on Strep-Tactin Sepharose (5 mg/ml) under gravity flow and physiological conditions (100 mM Tris-Cl, pH 8.0). The purification is documented on a Coomassie stained SDS gel where samples from the crude lysate (lane 1), from the flow through (lane 2), and from the elution with 2.5 mM desthiobiotin (lane 3) had been applied. The wild type enzyme is shown to be over 99 % pure (lane 3 contains 50 µg protein vs 0.5 µg protein per band in the molecular size standard). The mutant - although expressed at low level only - could be obtained at high purity under the same conditions.

1: Sample of crude lysate after cytosolic expression with pASK-IBA3
2: Sample of flow through during chromatography
3: Sample after the addition of 2.5 mM desthiobiotin
M: Molecular size standard (kDa)

Specifications of Strep-Tactin Sepharose resin

Strep-Tactin concentration	5 mg/ml sedimented resin
Binding capacity	1 ml sedimented resin is useful for the one-step purification of 50 to 100 nmol recombinant protein (up to 3 mg in case of a 30 KDa protein)
Support	Sepharose 4FF, 4 % agarose
Bead size	45 - 165 µm
Linear flow rate	gravity flow
Exclusion limit	3 x 10⁷ Da
Form	50 % suspension in 100 mM Tris/HCl pH 8.0, 1 mM EDTA, 150 mM NaCl
Stability	at least 6 months after shipment
Storage	4 °C, DO NOT FREEZE
Shipment	RT

View more information on the Strep-tag purification system and further specifications of column formats.

For Buffers & reagents pease click here

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Order information
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product	amount	cat. no.	order & prices
Gravity flow Strep-Tactin Sepharose mini-columns (0.2 ml bed volume)	5 columns	2-1202-550
Gravity flow Strep-Tactin Sepharose column (1 ml bed volume)	1 column	2-1202-001
	5 columns	2-1202-050
Gravity flow Strep-Tactin Sepharose column (5 ml bed volume)	1 column	2-1202-051
Gravity flow Strep-Tactin Sepharose column (10 ml bed volume)	1 column	2-1202-101
Strep-Tactin Sepharose (50 % suspension)	4 ml	2-1201-002
	20 ml	2-1201-010
	50 ml	2-1201-025
	200 ml	2-1201-100
	1000 ml	2-1201-500



		Strep-Tactin^® Sepharose^®

Strep-Tactin® Sepharose®

for purification of Strep-tag® II proteins by gravity-flow column chromatography

Strep-Tactin^® Sepharose^®

for purification of Strep-tag^® II proteins by gravity-flow column chromatography