Insights from the Blog
There are different digital imaging technologies available for Western blot protein detection. How do you choose between using a CCD camera and a laser scanner?Continue reading
The polyhistidine-tag is the most widely used affinity tag in research involving recombinant proteins. It is small, and hence, the first choice in many protein purification workflows. Purification of his-tagged proteins is often straightforward using an immobilized metal ion affinity chromatography (IMAC) resin. Still, low or no recovery of your target protein can occur.
His-tagged protein purification is popular, but doing it well is a bit of an art. I recently attended a colleague’s webinar and wanted to share some of her tips for a successful outcome. I also recommend that you listen to her presentation and the live question and answer session.
Based on responses from happy scientists, we are sharing these automated methods for removing an affinity tag and the protease. The tag is cleaved while the protein is still bound to the affinity resin. The next step depends on the protease. Thrombin or Factor Xa is removed by connecting a second (Benzamidine) column. Tagged proteases are removed on the same column used for capture.