t7 tag affinity purification kit  (Millipore)

Journal: PLoS ONE

Article Title: The plant defense and pathogen counterdefense mediated by Hevea brasiliensis serine protease HbSPA and Phytophthora palmivora extracellular protease inhibitor PpEPI10

doi: 10.1371/journal.pone.0175795

Figure Lengend Snippet: The T7 antibody covalently linked agarose beads were incubated with the purified His-T7-tagged PpEPI10 (+ rPpEPI10) or buffer (- rPpEPI10). The mixture was added to buffer (- HbSPA) or the intercellular fluids isolated from N . benthamiana leaves expressing HbSPA-His protein (+ HbSPA). After washing, the eluates were analyzed by Western blot with HRP conjugated anti-His monoclonal antibody. Lane M represents the protein standard. The numbers on the left indicate the size of molecular weight markers.

Article Snippet: Co-immunoprecipitation (co-IP) was performed on the intercellular fluid containing the His-tagged HbSPA and the purified His-T7-tagged PpEPI10 (rPpEPI10) using the T7.tag ® Affinity Purification Kit (Novagen, Merck Millipore, MA, USA) following the manufacturer’s instructions, with minor modifications.

Techniques: Incubation, Purification, Isolation, Expressing, Western Blot, Molecular Weight