PolyHunter

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  • 19103
Solubilization of membrane protein using synthetic copolymer nanodiscs has a lot of... more
Product information "PolyHunter"
Solubilization of membrane protein using synthetic copolymer nanodiscs has a lot of advantages over traditional membrane protein solubilization methods.
Cube Biotech's PolyHunter Agarose Resins and MagBeads solve one of the few remaining issues with this product. After solubilization, some remaining free copolymer molecules can interfere with the subsequent affinity chromatography. Our PolyHunter products can deplete up to 99% of these interfering copolymer molecules from your solubilizate.

PolyHunter Agarose Resin and PolyHunter MagBeads are compatible with all nanodisc copolymers that CubeBiotech offers. Therefore it is perfectly suited to be purchased in conjunction with our Synthetic nanodisc Screening Kit Maxi.

Note: PolyHunter Products are reusable up to 5 times. The corresponding regeneration procedure can be found in the matching protocol PDF files.
Schematic depiction on how PolyHunter Agarose worksFigure 1: Schematic depiction of a gravity flow column that depletes a copolymer solubilizate from its remaining free copolymer molecules.

Application Data

PolyHunter depletion efficencies compared betweeen CopolymersFigure 2: Average copolymer depletion efficiencies of PolyHunter beads of the four copolymer classes that Cube Biotech offers. Note that the depletion efficiency of DIBMA is lower that the rest. You can solve this problem by performing the PolyHunter protocols twice.
Example Protein Purification between PolyHunter treated solubiluzate and one withoutFigure 3: Affinity chromatography (AC) of a 9-transmembrane-domain protein. Each step of the protein purification procedure was performed with and without PolyHunter treatment.
S: Original solubilizates, both of them containing the same amount of protein
FT: Flow-through after the binding step of the AC. The Polyhunter treated flow-through contains less protein and therefore more has successfully bound to the affinity beads.
W: Wash fractions (near invisible), in both cases next to no protein was eluted during the washing steps.
E: Elution fractions, PolyHunter treated sample shows a higher protein yield.
The need for repeated cycles when using Polyhunter Magbeads (Step 3-6)Figure 4: Copolymer depletion efficiency per cycle when using PolyHunter Magbeads. The protocol of the PolyHunter MagBeads requires performing stepsa 3-6 three times in a row. These depletion schemes are the reason behind it.
AASTY 6-55 depletion efficiency by PolyHunter productsFigure 5: Depletion efficiency of PolyHunter products of AASTY 6-55. In this depletion, a near 100% efficiency was reached.
Repeated Depletion efficiencies of AASTY 6-50 after several regeneration steps of PolyHunter BeadsFigure 6: Depletion efficiencies of AASTY 6-50 measured at wavelength 260nm. Note that even after 5 regenerations (see protocol) of the PolyHunter beads the depletion efficiency is nearly identical, if not greater than the first "fresh" run.
Example Protein Purification between PolyHunter treated solubiluzate and one withoutFigure 7: Depletion efficiencies of AASTY 11-50 measured at wavelength 260nm. Note that even after 5 regenerations (see protocol) of the PolyHunter beads the depletion efficiency is nearly identical, if not greater than the first "fresh" run.
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