StrepTactin® Agarose | For Strep® tag purification | Cube Biotech

StrepTactin Agarose


Agarose purification resin
The Strep-tag® purification system is based on the highly selective binding of engineered streptavidin, called Strep-Tactin, to Strep-tag II fusion proteins. Cube Biotech offers a high-quality affinity resin for strep-tagged proteins, with maximum binding capacity. PureCube HiCap StrepTactin Agarose is delivered as a 50% suspension. For purification from cell culture supernatants or for pull-down experiments, PureCube HiCap StrepTactin® MagBeads are recommended. For detection of strep-tagged proteins in Western Blots, specific antibodies and antibody conjugates are available.

PureCube HiCap StrepTactin Agarose provides:

  • Binding capacity of up to 5 mg protein per mL resin
  • Stable in the presence of a wide range of additives
  • High-quality performance for batch or gravity flow purification
  • Also available as MagBeads and prepacked FPLC columns

Features

Usage Specific binding and purification of 6x his-tagged proteins
Specifity Affinity to His-tagged proteins
Binding capacity >5 mg/mL
Compatibility 2% Tween 20, 2% Triton X-100, 2% IGEPAL® 630/Nonidet P40, n-Octyl-beta-D-glucopyranoside, 0,2% n-Nonyl-beta-D-gluyopyranoside, 0,35% N-Decyl-beta-D-maltoside, 2% Lauryl-sacrosine, 0,1% SDS, 0,3% CHAPS, 1M Guanidine Hcl, 1mM PMSF, 10% Ethanol, 5M NaCl, 2M (NH4)SO4, 1M CaCl2, 25% Glycerol
Filling quantity Delivered as a 50 % suspension
Bead size 100 μm, to obtain flow rates up to 6 mL/min
Bead Ligand StrepTactin® Agarose
Required equipment
 
  • Lysis Buffer
  • Wash Buffer
  • Elution Buffer
  • Ice bath
  • Refrigerated centrifuge for 50 mL tube (min 10,000 x g)
  • 50 mL centrifuge tube
  • Micropipettor and Micropipetting tips
  • Disposable gravity flow columns with capped bottom outlet, 2 ml
  •  pH meter
  •  Optional: 15 mL conical propylene tubes (e.g. Falcon)
  • End-over-end shaker
  • SDS-PAGE buffers, reagents and equipment Optional: Western Blot reagents and equipment

Applications

All protocols and buffer compositions are also avaible as PDF-Files on the Protocols & Datasheets page.
   
A.Protein purification protocol
 
 
  1. . Thaw the E. coli cell pellets corresponding to 200 mL bacterial culture on ice for 15 min. Optional: Freezing the cell pellet at –20˚C for 30 min prior to incubation at room temperature improves lysis by lysozyme.
  2. Resuspend the cell pellet in 10 mL Lysis Buffer, and pour it into a 50 mL conical centrifuge tube.
  3. If the solution is very viscous, add 3 units Benzonase® per mL E.coli culture volume to the lysis buffer. Alternatively or additionally, sonicate the lysate to improve cell disruption.
  4. Incubate on an end-over-end shaker at 4°C for 1 h.
  5. . Centrifuge the lysate for 30 min at 10,000 x g and 4˚C. Carefully collect the supernatant without touching the pellet. Note: The supernatant contains the soluble proteins and is the cleared lysate fraction. We recommend to take aliquots of all fractions for SDS-PAGE analysis.
  6. Resuspend the PureCube HiCap StrepTactin Agarose by inverting the bottle until the suspension is homogeneous. Transfer 1 mL of the 50% suspension (corresponding to 500 µL bed volume) to a 15 mL conical centrifuge tube. Allow the resin to settle by gravity and remove the supernatant. Tip: Alternatively, resin equilibration can be performed directly in the disposable gravity flow column.
  7. Add 3 mL Wash Buffer and gently resuspend the slurry to equilibrate the resin. Allow the resin to settle by gravity and remove the supernatant. Repeat this step twice.
  8. Add 10 mL cleared lysate to the equilibrated PureCube HiCap StrepTactin Agarose resin and incubate at 4˚C for 1 h on an end-over-end shaker.
  9. Transfer the binding suspension to a disposable gravity flow column with a capped bottom outlet. Use Wash Buffer to rinse the centrifuge tube and remove resin adhered to the wall. Tip: Alternatively, batch binding can be performed directly in a gravity flow column with closed bottom and top outlets.
  10. Remove the bottom cap of the column and collect the flowthrough. This is the flow-through fraction.
  11. Wash the column with 1 mL Wash Buffer. Repeat the washing step at least 3 times. These are the wash fractions
  12. Elute the Strep-tagged protein 5 times using 250 µL Elution Buffer. Collect each eluate in a separate tube and determine the protein concentration of each fraction. These are the elution fractions. Optional: Incubate the resin for 15 min in Elution Buffer before collecting the eluate to increase protein yields.
  13. Analyze all fractions by SDS-PAGE. Note: Do not boil membrane proteins. Instead, incubate samples at 46˚C for 30 min in preparation for SDS-PAGE analysis.
  14. . Optional: Perform Western Blot using Strep Antibody.
StrepTactin® and Strep-tag® are trademarks of IBA GmbH. PureCube StrepTactin Agarose is manufactured by IBA GmbH under German Patent Application No. 42 37 113.9 entitled "Fusion peptides with binding activity for streptavidin".
Agarose purification resin
The Strep-tag® purification system is based on the highly selective binding of engineered streptavidin, called Strep-Tactin, to Strep-tag II fusion proteins. Cube Biotech offers a high-quality affinity resin for strep-tagged proteins, with maximum binding capacity. PureCube HiCap StrepTactin Agarose is delivered as a 50% suspension. For purification from cell culture supernatants or for pull-down experiments, PureCube HiCap StrepTactin® MagBeads are recommended. For detection of strep-tagged proteins in Western Blots, specific antibodies and antibody conjugates are available.

PureCube HiCap StrepTactin Agarose provides:

  • Binding capacity of up to 5 mg protein per mL resin
  • Stable in the presence of a wide range of additives
  • High-quality performance for batch or gravity flow purification
  • Also available as MagBeads and prepacked FPLC columns

Features

Usage Specific binding and purification of 6x his-tagged proteins
Specifity Affinity to His-tagged proteins
Binding capacity >5 mg/mL
Compatibility 2% Tween 20, 2% Triton X-100, 2% IGEPAL® 630/Nonidet P40, n-Octyl-beta-D-glucopyranoside, 0,2% n-Nonyl-beta-D-gluyopyranoside, 0,35% N-Decyl-beta-D-maltoside, 2% Lauryl-sacrosine, 0,1% SDS, 0,3% CHAPS, 1M Guanidine Hcl, 1mM PMSF, 10% Ethanol, 5M NaCl, 2M (NH4)SO4, 1M CaCl2, 25% Glycerol
Filling quantity Delivered as a 50 % suspension
Bead size 100 μm, to obtain flow rates up to 6 mL/min
Bead Ligand StrepTactin® Agarose
Required equipment
 
  • Lysis Buffer
  • Wash Buffer
  • Elution Buffer
  • Ice bath
  • Refrigerated centrifuge for 50 mL tube (min 10,000 x g)
  • 50 mL centrifuge tube
  • Micropipettor and Micropipetting tips
  • Disposable gravity flow columns with capped bottom outlet, 2 ml
  •  pH meter
  •  Optional: 15 mL conical propylene tubes (e.g. Falcon)
  • End-over-end shaker
  • SDS-PAGE buffers, reagents and equipment Optional: Western Blot reagents and equipment

Applications

All protocols and buffer compositions are also avaible as PDF-Files on the Protocols & Datasheets page.
   
A.Protein purification protocol
 
 
  1. . Thaw the E. coli cell pellets corresponding to 200 mL bacterial culture on ice for 15 min. Optional: Freezing the cell pellet at –20˚C for 30 min prior to incubation at room temperature improves lysis by lysozyme.
  2. Resuspend the cell pellet in 10 mL Lysis Buffer, and pour it into a 50 mL conical centrifuge tube.
  3. If the solution is very viscous, add 3 units Benzonase® per mL E.coli culture volume to the lysis buffer. Alternatively or additionally, sonicate the lysate to improve cell disruption.
  4. Incubate on an end-over-end shaker at 4°C for 1 h.
  5. . Centrifuge the lysate for 30 min at 10,000 x g and 4˚C. Carefully collect the supernatant without touching the pellet. Note: The supernatant contains the soluble proteins and is the cleared lysate fraction. We recommend to take aliquots of all fractions for SDS-PAGE analysis.
  6. Resuspend the PureCube HiCap StrepTactin Agarose by inverting the bottle until the suspension is homogeneous. Transfer 1 mL of the 50% suspension (corresponding to 500 µL bed volume) to a 15 mL conical centrifuge tube. Allow the resin to settle by gravity and remove the supernatant. Tip: Alternatively, resin equilibration can be performed directly in the disposable gravity flow column.
  7. Add 3 mL Wash Buffer and gently resuspend the slurry to equilibrate the resin. Allow the resin to settle by gravity and remove the supernatant. Repeat this step twice.
  8. Add 10 mL cleared lysate to the equilibrated PureCube HiCap StrepTactin Agarose resin and incubate at 4˚C for 1 h on an end-over-end shaker.
  9. Transfer the binding suspension to a disposable gravity flow column with a capped bottom outlet. Use Wash Buffer to rinse the centrifuge tube and remove resin adhered to the wall. Tip: Alternatively, batch binding can be performed directly in a gravity flow column with closed bottom and top outlets.
  10. Remove the bottom cap of the column and collect the flowthrough. This is the flow-through fraction.
  11. Wash the column with 1 mL Wash Buffer. Repeat the washing step at least 3 times. These are the wash fractions
  12. Elute the Strep-tagged protein 5 times using 250 µL Elution Buffer. Collect each eluate in a separate tube and determine the protein concentration of each fraction. These are the elution fractions. Optional: Incubate the resin for 15 min in Elution Buffer before collecting the eluate to increase protein yields.
  13. Analyze all fractions by SDS-PAGE. Note: Do not boil membrane proteins. Instead, incubate samples at 46˚C for 30 min in preparation for SDS-PAGE analysis.
  14. . Optional: Perform Western Blot using Strep Antibody.
StrepTactin® and Strep-tag® are trademarks of IBA GmbH. PureCube StrepTactin Agarose is manufactured by IBA GmbH under German Patent Application No. 42 37 113.9 entitled "Fusion peptides with binding activity for streptavidin".
Top seller
StrepTactin Agarose
No results were found for the filter!
PureCube HiCap StrepTactin Agarose PureCube HiCap StrepTactin Agarose
2 ml 50% High Capacity StrepTactin Agarose
Article number: 34101
Sales price: From €67.00 *
Viewed