Cat #: 11-718
Zymo Research P2004 Strep-Spin Protein Miniprep Kit, Zymo Research, 10 Preps/Unit
Cat #: 11-718
Zymo Research P2004 Strep-Spin Protein Miniprep Kit, Zymo Research, 10 Preps/Unit
Zymo Research
10 Preps/Unit
Brand: Zymo Research- Fast: Purify Strep-tagged proteins from cell lysates in less than 7 minutes
- Simple: Prepare pure protein for small-scale studies using a spin-column
- Convenient: No special instrumentation needed other than a benchtop microcentrifuge
$109.90
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Zymo Research
10 Preps/Unit
Brand: Zymo Research- Fast: Purify Strep-tagged proteins from cell lysates in less than 7 minutes
- Simple: Prepare pure protein for small-scale studies using a spin-column
- Convenient: No special instrumentation needed other than a benchtop microcentrifuge
The Strep-Spin Protein Miniprep Kit™ provides a fast spin-column based purification technology for Strep-tagged proteins. Up to 450µg of Strep-tagged protein can be purified in 7 minutes and eluted in Strep-Elution Buffer. The purified protein is ultra-pure and can be used directly for enzymatic assays, protein biochemical analyses, SDS-PAGE and other sensitive applications. The straightforward spin, wash, and elute protocol dramatically simplifies protein purification and allows the end user to get results in minutes, not hours.
| Brand | Zymo Research Corporation |
|---|---|
| Affinity Matrix | Strep-Tactin® XT Superflow® 50% |
| Binding Capacity | 100µl of Strep-Tactin® XT Superflow® 50% suspension can bind 15nmol biotin (approx. 450µg of a 30 kDa Twin-Strep-tag® protein). |
| Elution Method | Biotin excess |
| Elution Volume | Recommended 3 x 200µl |
| Principle of Technology | The Strep-Spin Protein Miniprep Kit is based on a novel StrepTactin® XT Superflow® resin which binds to Twin Strep-tag® with very high affinity. This tag contains two Strep-tag® II peptides connected via a Glycine/Serine linker sequence. Hence this kit is optimized for the purification of Twin-Step®-tagged proteins; however it will also work efficiently for proteins with single Strep-tags. |
| Processing Time | 7 minutes |
| Product Storage | Please store the Strep-Tactin® XT Superflow® 50% suspension, Strep-Wash Buffer, and Strep-Elution Bufer at 4°C. The other components can be stored at room temperature. |
| Protein Purity | Electrophoretically pure. Purified high-quality protein is suitable for enzyme kinetics, protein biochemical analyses, SDS-PAGE, and other applications. |
| Required Equipment | Microcentrifuge |
| Sample Type | Cell lysates or other complex protein mixtures containing Strep-tagged proteins. |
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Q1: Which Step-Tag amino acid sequences are recommended?
Strep-tag® II: WSHPQFEK Twin-Strep-Tag® (Strep-tag III): WSHPQFEKGGGSGGGSGGWSHPQFE
Membrane proteins can be purified after solubilization in a nonionic detergent. Concentrations of up to 2% of Triton® or TWEEN® can be present in the loaded sample.
No, this will prevent the protein from binding to the matrix. To remove biotin from the sample we recommend to add stoichiometric amounts of Avidin prior to using the kit. Avidin blocks the biotin but does not bind to the Strep-tag. Alternatively, dialysis could be conducted.
The pH value of the loaded sample should be greater then 7.0.
The sample may contain up to: Reducing Agents </= 50 mM beta-mercaptoethanol Non-Ionic Detergents </= 2% Nonidet® P-40 </= 2% Triton® X-100 </= 2% Tween®-20 Ionic Detergents </= 0.1% SDS (Sodium-N-dodecyl sulfate) Denaturing Reagents </= 8M Urea Chemicals and other Reagents </= 50mM EDTA </= 10% (v/v) Ethanol </= 25% (v/v) Glycerol </= 250mM Imidazole </= 1M MgCl2 </= 5M NaCl
For protein purification from E. coli lysates: 1. Harvest & pellet 10ml of E. coli culture. 2. Resuspend in 1-2ml of Lysis Buffer (e.g. 300mM NaCl,1% NP-40 in PBS, 1mg/ml Lysozyme, 1 x Protease Inhibitor, 15U/ml Benzonase®). 3. Incubate for 30 minutes at room temperature. 4. Spin at >/= 12,000 x g at 4°C for 5 minutes. 5. Use the supernatant for the Strep-Spin Protein Miniprep Kit™ protocol.
Check your buffers for signs of contamination and check the pH of the buffers. - Increase centrifugation time and speed. Ensure that the Strep-Tactin® XT Superflow® drains completely after each spin (some older centrifuge models may require a longer centrifugation time). - If the problem persists, additional wash steps can be added to the purification protocol.
Smaller elution volumes are possible and may yield more concentrated protein, but the elution efficiency may be compromised.
It will work without the equilibration step, however, for optimal performance we recommend including it.
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