Tandem Affinity Purification (TAP)-MS
Tandem affinity purification (TAP) is a universal two-step affinity purification protocol used to separate TAP-labeled proteins and related proteins. The principles of TAP and COIP technology for screening interacting proteins are similar. The difference is that the TAP technology puts two purification tags on the bait protein. Two rounds of purification of the protein complex (tandem affinity purification) are carried out using the tags to reduce the non-specific proteins in the eluate to a lower level. Combined with the LC-MS/MS technology, the protein types in the eluate of the experimental group and the negative control group were identified, so as to find the unique protein in the experimental group, that is, the possible interacting protein.
The TAP-MS technical process is as follows:
- Construct a vector containing the gene of interest
- The vector transfects cells and expresses the fusion protein
- Cell lysis to extract total protein
- Purification and elution
- Mass spectrometer analysis and identification of proteins that interact with bait proteins
Profacgen provides the following services for you:
- Construction of bait protein eukaryotic expression vector or lentiviral vector fused with Flag-HA
- Transient vector transfection of target cells or packaged lentivirus infection of target cells
- TAP purifies the interacting protein and reduces the number of non-specific binding proteins. We provide including but not limited to protein molecular weight and isoelectric point determination, protein subunit analysis, specific protein detection, etc.
- The interacting proteins were identified by mass spectrometry and proteomics was analyzed on a large scale.
The TAP-MS service provided by Profacgen is based on a two-step purification system, which significantly reduces the binding rate of non-specific proteins, while the complex is relatively stable, and the results are accurate and reliable.
Profacgen has an experienced team that provides a variety of TAP/MS services. Please contact us for specific project implementation content and time. Profacgen will be your trusted partner.