Summary
Zymo-Spin P1 Column His-tag Purification Kit
Protein kept on ice during procedure; centrifuge in cold room used.
- Vortexed His-Affinity Gel briefly.
- Transferred 250μL of His-Affinity Gel to the Zymo-Spin P1 column.
- Placed column in 2mL collection tube.
- Centrifuged for 20 seconds at max rpm.
- Discarded flow-through.
- Added 300μL PcTF in protein extraction buffer to column.
- End-over-end mixing for two minutes to resuspend gel and allow protein to bind to resin.
- Centrifuged for 20 seconds at max rpm.
- Discarded flow-through.
- Added 250μL His-Wash Buffer and mixed to resuspend gel.
- Centrifuged for 20 seconds at max rpm.
- Discarded flow-through.
- Repeated wash step once.
- Transferred column into a labeled 1.7mL microcentrifuge tube.
- Added 150μL His-Elution BUffer and resuspended gel via end-over-end mixing for one minute.
- Centrifuged for 20 seconds at max rpm.
- Discarded column and stored purified protein at -20°C.
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