Visual protocol: Capturem his-tagged maxiprep kit

This protocol provides simple, rapid purification of his-tagged proteins from up to 25 ml of clarified lysate from mammalian or bacterial cell samples using the Capturem His-Tagged Purification Maxiprep Kit (Cat. # 635713). These columns are suitable for use under native or denaturing conditions, and in the presence of additives such as DTT (up to 10 mM), βME (up to 30 mM), TCEP (up to 5 mM), EDTA (up to 10 mM), or glycerol.

	Pre-purification filtration Load 2–25 ml cleared lysate onto the Capturem Maxiprep Filter (purple insert) Spin for 3 min Discard filter
	Save filtered lysate for Step 2
	Step 1: Equilibrate Add 6 ml xTractor Buffer to the Capturem Maxiprep Nickel Column (clear insert) Spin for 3 min Discard flowthrough
	Step 2: Bind Load 2–25 ml filtered lysate from the pre-purification filtration step onto the Capturem Maxiprep Nickel Column Spin for 3 min Transfer column to a new collection tube and save flowthrough
	Step 3: Wash Add 6 ml Wash Buffer* to the Capturem Maxiprep Nickel Column Spin for 3 min Transfer column to a new collection tube and save flowthrough *Optimize washing conditions by adding 200–800 µl Elution Buffer to change the imidazole concentration.
	Step 4: Elute Add 1 ml Elution Buffer to the Capturem Maxiprep Nickel Column Spin for 3 min Collection tube contains your eluted tagged protein

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Visual protocol: Capturem his-tagged maxiprep kit

Pre-purification filtration

Step 1: Equilibrate

Step 2: Bind

Step 3: Wash

Step 4: Elute