HIS-Tag Protocol

Purification & Detection

HIS-Tag Immuno-Precipitation Protocol using Magnetic Agarose Beads

Product No. H9914

Prior to use
Ni magnetic beads supplied in ethanol are repeatedly exchanged with wash buffer and used at a final concentration of 10% (v/v).

Step 1: Tranfer beads to cell lysate
Add 30 µl of Ni magnetic beads to soluble cell lysate and mix for 15 min at RT.

Step 2: Remove lysate
Place 96-deep well block onto a 96-well magnet for 5 min. to separate the magnetic beads in solution. Remove the lysate and discard.

Step 3: Wash beads
Add 1 ml of wash buffer to 96-deep well block, shake for 5 min. to resspend Ni magnetic beads, place again on the 96-well magnet and remove buffer. Repeat washing step.

Step 4: Add elution buffer
Add 60 µl elution buffer and shake for 3 min. Spin plate in centrifuge to ensure that all beads come in contact with the elution buffer.

Step 5: Transfer elution proten
Place 96-deep well block on 96-well magnet to separate the magnetic beads and elution buffer. Transfer 60 µl eluted protein to a 96-well microtiter plate.