Haynes Lab:Notebook/Engineering PC-TFs/2014/09/25: Difference between revisions
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==Summary== | ==Summary== | ||
* | * Purified PcTF protein. | ||
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'''Zymo-Spin P1 Column His-tag Purification Kit'''<br> | |||
Protein kept on ice during procedure; centrifuge in cold room used.<br> | |||
*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. | |||
Revision as of 11:20, 27 September 2014
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Summary
Zymo-Spin P1 Column His-tag Purification Kit
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