The GST (Glutathione S transferase) pull-down has proven to be a powerful technique for studying protein-protein interactions, allowing the identification of unknown proteins that interact with known proteins and confirming suspected interactions, as well as identifying the presence of interactions between two known proteins.
Principle of GST Pull-down
Mechanistically, the probe protein is fused with GST by recombinant technology, and the fusion protein can then be bound to GTH (Glutathione) on the solid support through GST due to affinity attraction. Therefore, proteins that interact with fusion proteins when passing through the column or mixed with the solid phase complex are adsorbed and separated.
GST-pull down mainly consists of the following three parts: (i) construction of prokaryotic expression vector with GST tag by recombinant technology; (ii) expression of the fusion protein with GST tag by prokaryotic expression system; (iii) obtaining high purity fusion protein via the GST affinity purification column and using the GST affinity purification column for protein-to-protein interaction detection; (iv) using WB and MS detection to verify the interaction between proteins or screen for target proteins.
Advantages and Disadvantages of GST Pull-down
| Advantages | Disadvantages |
|---|---|
| Easy to operate can directly detect interactions | Requires sufficient preparation of soluble recombinant proteins Only suitable for the detection of stable or strong interactions Protein interactions may interfere with endogenous proteins |
GST Pull-down Applications
Demonstrate possible interactions between two known proteins
Discover unknown molecules that interact with known proteins
Our GST Pull-down Services
As a leading proteomic technology service company, Creative Proteomics has many years of experience in mass spectrometry-based protein interaction analysis. Creative Proteomics provides efficient strategies for protein expression and purification, as well as optimal optimization solutions for GST pull-down. Featuring strict quality control and turnaround time, we provide customers with customizable one-stop services covering every step of your project, from gene synthesis to data reporting, hence maximizing your cost-efficiency.
| Service | Provides | Deliverables |
|---|---|---|
| GST-protein preparation (optional) GST pull-down | Purified protein Plant and animal tissues Cell lysis solution | Experimental reports |
Workflow of GST Pull-down
Our Advantages
- Setting up multiple control groups that fully exclude interference and ensure reliable results
- Repeated experiments (twice) to exclude random factor influences
- Complete downstream package for further quantitative analysis using the BLI technology
- Utilizing the silver staining technology with 100 times higher sensitivity than Komas Brilliant Blue to ensure clearer and better SDS-PAGE electrophoresis results
- One-stop service from GST protein preparation to protein interaction detection
- Higher affinity and specificity
Protein-protein interaction (PPI) is the basis of many life activities, so the research enthusiasm for PPI has not diminished. For over a decade, Creative Proteomics has been devoted to establishing innovative and advanced PPI analysis platforms, covering numerous excellent technologies and assays that can help customers in studying the PPI in vivo, in vitro, and in silico. We are honored to become your competent research assistant.