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Products >  Protein_Expression_and_Purification >  Protein_Learning_Center >  Learning_Resources >  His-Tagged_Miniprep_Visual_Protocol

Visual Protocol: His-Tagged Purification Miniprep Kit

VISUAL PROTOCOL

Capturem His-Tagged Purification Miniprep Kit

This protocol provides simple, rapid purification of his-tagged proteins in up to 800 µl of clarified lysate from mammalian or bacterial cell samples using the Capturem His-Tagged Purification Miniprep Kit (Cat. No. 635710). The columns are suitable for use under native or denaturing conditions, 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.

Equilibrate

400 µl

  • Add 400 µl xTractor Buffer
  • Spin for 1 min
  • Discard flowthrough

Bind

up to 800 µl of cleared lysate

  • Load 200–800 µl cleared llysate
  • Spin for 1 min
  • Transfer column to new collection tube and save flowthrough

Wash

300 µl

  • Add 300 µl Wash Buffer*
  • Spin for 1 min
  • Transfer column to new collection tube and save flowthrough

*Optimize washing conditions by adding 5–30 µl Elution Buffer to change the imidazole concentration.

Elute

100–300 µl

  • Add 100–300 µl Elution Buffer
  • Spin for 1 min
  • Collection tube contains your eluted tagged protein
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