GST pull-down assay is an in vitro assay that effectively validates the yeast two-hybrid system. The target protein-GST fusion protein is affinity-cured on the glutathione affinity resin as a support for affinity with the target protein, acting as a "bait protein", and the target protein solution is passed through the column, and the interaction with the target protein can be captured therefrom. "Capture protein (target protein)", the eluted conjugate was separated by SDS-PAGE electrophoresis, and finally western blot was performed to confirm the interaction between the two proteins.
Both “bait protein” and “capture protein” can be obtained by cell lysate, purified protein, expression system and in vitro transcription translation system. This method is simple and easy to operate.
1. Construct the purified target protein and related bait protein or cell lysate supernatant containing the bait protein .
2. In vitro GST pull-down experiment.
1. Interact protein name, Gene ID, species information.
2. Decoy protein and capture protein expression vector.
3. Detection of primary antibodies (also available from AtaGenix).
1. PAGE electrophoresis results.
2.Western blot original picture and identification results.
3. Experimental report
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In vitro GST pull down (Prokaryotic target protein and bait protein)
In vitro GST pull down (Prokaryotic target protein and cell lysate supernatant)
In vitro GST pull down (Eukaryotic target protein and cell lysate supernatant)
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