Strep-tag®-protein immobilization

Immobilization of proteins can be useful for the following applications:

Antibody or serum screening
Diagnostic assays
Expression cloning
Protein interaction studies
Screening of engineered enzymes
Drug screening

IBA products

Strep-Tactin® coated microplates

and

for

Oriented immobilization of recombinant proteins with N-terminal, C-terminal or internal Strep-tag® II
Effective screening procedures
Minimal non-specific binding
High and constant activity
Minimal coefficients of variation (cv)

Specifications

Specifications of Strep-Tactin® coated microplates

Strep-tag® fusion protein binding capacity	100 ng Strep-tag® II fusion protein/well
Intra- and inter-assay variance (cv)	< 1.6 % and < 3.0 %, respectively
Max. volume per well	300 µl
Optimal pH	8.0
Stability	> 1 year at 4 °C
Storage	4 °C
Shipment	RT

Specifications of StrepMAB-Immo

Host animal/Subclass	Mouse/IgG1
Specifity	Nearly irreversible binding of C- or N-terminally fused Strep-tag® II, which is N-terminally extended by "SerAla" ** like recombinant protein - SA-WSHPQFEK or SA-WSHPQFEK - recombinant protein. Proteins containing Strep-tag® II with N-terminal sequences other than SerAla are bound with reduced affinity in the cases tested so far.
Form binding capacity	lyophilized from NH₄HCO₃with 10 mg sucrose per mg antibody (determined for bacterial alkaline phosphatase)
Stability	24 months after shipment
Storage	-20 °C
Shipment	RT

**pASK-/pEXPR- and pPR-IBA vectors with N-term One-/Strep-tag® II do NOT have "SerAla"-Linker but the
StarGate® vectors (carrying Strep-tag® II and One-STrEP-tag) are equipped with the SA-Linker.

Specifications of StrepMAB-Immo coated microplates

Plate type	12 x 8-well strips and frame, transparent, flat bottom
StrepMAB-Immo coated area	100 µl
Blocked area	300 µl
Strep-tag® II fusion protein	~3 pmol Strep-tag® II fusion protein per 100 µl
binding capacity	~3 pmol Strep-tag® II fusion protein per 100 µl incubation volume determined for bacterial alkaline phosphatase
Max. volume per well	300 µ
Specificity	Nearly irreversible binding (K_D StrepMAB Immo: SerAla- Strep-tag® II ~ 1 pM) of C- and N-terminally fused Strep-tag® II with SerAla linker seq.- SerAlaTrpSerHisPro GlnPheGluLys). The Strep*-tag® will be linked automatically to the C-terminus of the recombinant protein via the SerAla dipeptide when the vectors for C-terminal fusion from IBA are used. Other linker sequences than SerAla lead to reduced affinity in the cases tested so far.
Recommended buffers	Binding: 25 mM Tris-HCl, 2 mM EDTA, 140 mM NaCl, pH 7.6 (optional pH 7.4-8.0) Washing: 0.05% Tween-20 in binding buffer or PBS
Stability	6 months after shipment
Storage	4 °C
Shipment	RT

Specifications of Gravity flow StrepMAB-Immo MacroPrep Columns

Specifications of Gravity flow StrepMAB-Immo MacroPrep Columns	Nearly irreversible binding of C- and N-terminally fused Strep-tag® II with SerAla linker (rec. protein seq.- SerAlaTrpSerHisProGlnPheGlu Lys). The Strep-tag® II will be linked automatically via the SerAla dipeptide to the C-terminus when the vectors for C-terminal fusion from IBA are used. Other linker sequences than SerAla lead to reduced affinity in the cases tested so far.
Support	MacroPrep® 50 (polymethacrylate)
Form	Pre-packed in 100 mM Tris/HCl pH 8.0, 1 mM EDTA, 150 mM NaCl
Strep-tag® II fusion protein binding activity	> 15 nmol/ml resin
Stability	6 months after shipment
Storage	4 °C
Shipment	RT

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