Product name: Strep-Tactin Sepharose FF


English name: Strep-Tactin NUPharose Fast Flow, Strep-Tactin NUPharose FF


Specifications: 20 ml, 100 ml, 1L, 1 ml pre-packed column, 5 ml pre-packed column, special specifications customized


Transportation: 2 ~ 25℃, normal pressure, avoid light


Storage: 20% ethanol, 2 ~ 8℃


Shelf life: 3 years






Related introduction:


Strep-Tactin Sepharose FF is a bio-affinity chromatography separation medium formed by bonding Strep-Tactin on Sepharose microspheres. It is mainly used to purify Strep II tagged proteins. The Strep II tag is a small tag of 8 amino acids (WSHPQFEK). Because the tag is small, only about 1 kDa, it generally does not affect the structure and function of the fusion protein. It is often used for the detection and purification of the fusion expressed protein.






The ligand Strep-Tactin of this product is a mutant of Streptavidin. Compared with streptavidin, Strep-Tactin has at least 10 times stronger affinity for the Strep II tag, and can be used in a mild manner. Binding and dissociation with Strep II fusion protein under the conditions of the same. Since Strep-Tacin is highly specific to the Strep II tag, a high-purity protein sample can generally be obtained in one step.






After the Strep II-tagged protein is bound to the gel, it can be eluted using the desthiobiotin competition method. This elution method is relatively mild and generally does not affect the properties of the protein. For example, Strep II-labeled protein can tolerate alkaline conditions and can also be eluted with alkaline solutions, such as 10 mM NaOH.


Strep-Tactin Sepharose FF can tolerate higher alkaline conditions and can be regenerated and cleaned with 0.5 M NaOH and to remove heat sources.






In addition, 2-(4-hydroxybenzoic acid) benzoic acid (HABA) can also be used for gel regeneration. Excess HABA can replace desulphurized biotin in a competitive manner. However, in a HABA-free buffer, Strep-Tactin and HABA Dissociation will occur, causing HABA to fall off and achieve regeneration.






Technical index:


Matrix


4% agarose gel beads


Ligand


Strep-Tactin


Ligand Density


≥5 mg/ml


Filler particle size


60 ~ 180 μm


Maximum flow rate


800 cm/h


Recommended flow rate


20 ~ 100 cm/h


pH stability


Short-term pH 2 ~ 13; long-term pH 4 ~ 11


Back pressure resistant


0.3 MPa


Capacity


≥6 mg/ml Strep II tagged protein






Instructions:


1 Recommended buffer


Purified Strep II tagged protein


Binding buffer: 100 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, pH 8.0;


 　　　　　　 or 20 mM NaH2PO4, 280 mM NaCl, 6 mM KCl, pH 7.4






Elution buffer: binding buffer + 2.5 mM desthiobiotin;


 　　　　　　 or 10 mM NaOH






Regeneration buffer: 0.5 M NaOH;


 　　　　　　 or binding buffer+1 mM HABA






2 Sample preparation


The sample on the column should be as consistent as possible with the binding buffer. The samples can usually be processed by dialysis, ultrafiltration, dilution and other methods. And before loading the column, the insoluble matter should be removed by 0.45 μm filter membrane or high-speed centrifugation.






3 Sample purification


3.1 Equilibrium: Take an appropriate amount of Strep-Tactin Sepharose FF into a suitable chromatography column, wash 5 column volumes with distilled water to remove the preservation solution, and then equilibrate 5 column volumes with binding buffer. The recommended flow rate is 100 cm /h.


3.2 Sample loading: Load the prepared sample on the column. The recommended flow rate is 20 ~ 100 cm/h. The flow rate can be selected according to the actual combination to obtain better results.


3.3 Re-equilibration: After loading the sample, equilibrate more than 10 column volumes with binding buffer, or equilibrate to the baseline to wash away impurities. The recommended flow rate is 100 cm/h.


3.4 Elution: Wash 10-20 column volumes with elution buffer. The recommended flow rate is 100 cm/h. The collected eluate should be adjusted to a stable range immediately, and the buffer should be replaced as needed.


3.5 NaOH regeneration: After eluting the target protein, wash the column with distilled water for 3 ~ 5 column volumes, and use 0.5 M NaOH to regenerate 3 ~ 5 column volumes, then wash with distilled water to neutrality, store the column with 20% ethanol, or proceed Next purification.


3.6 HABA regeneration: If the target protein is eluted with desulphur biotin, it can also be regenerated with HABA buffer. Generally, wash 15 column volumes with HABA binding buffer, then wash 30 column volumes with binding buffer, and then use 20 Save the column in% ethanol, or proceed to the next step of purification. After HABA is loaded on the column, the gel color will change to reddish orange, and will return to normal white after the binding buffer is balanced. This regeneration method generally uses a larger volume.

Warm tips: Suzhou Beike nano products are only used for scientific research, not for human body,different batches of products have different specifications and performance

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