NTA versus IDA: what's the difference?:
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Nickel-NTA MagBeads:
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Indigo MagBeads:
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MagBead Separator:
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Wash Buffer and Elution Buffer:
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MagBeads Purification Steps:
Step 1:
The usual starting point of a MagBead protein purification is a cell lysate in which the proteins of interest are mixed together with every other proteins the expression cells had produced simultaneously.
Step 2:
Remove the storage buffer from the magnetic beads and wash two times with wash buffer. Add now an appropriate amount of magnetic beads to the lysate. We suggest that you use sterile pipette tips for this.
Step 3:
Shaking of the MagBead/Protein mixture. We suggest automated mixing with a slower but steady speed. No vortexing! A duration of around 2 hours at 4 °C is what we see as sufficient.
Step 4:
Now, almost every protein of interest should have bound to a MagBead.
Step 5:
Apply a magnetic force to your magnetic beads to separate them and in turn the protein of interest from the rest of the lysate. For micro centrifuge tube we suggest our practical MagBead separator for this.
Step 6:
In the final step the rest of the cell lysate that did not bind to the magnetic beads will be removed using again sterile peptide tips. Keep the magnetic force active to avoid accidental removal of some magnetic beads. Now wash the bound proteins and elute them with elution buffer.
Optional:
Additional washing steps will increase the purity of your protein. Different types of MagBeads can require different kinds of washing buffers. To see how the corresponding washing buffer for your MagBeads is prepared, consult our protocols & datasheets page.
You will also find the necessary information to elute the proteins of interest from the magnetic beads later on if this is necessary for you. Feel free to contact us in case you have any further questions.
Find a comprehensive guide at:
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7 сен 2024