A new affinity tag for the purification of recombinant proteins termed the ‘Dac-tag' has been developed by Axel Knebel who is the head of the Protein Production and Assay Development team at SCILLS. Together with the MRC and SCILLS cloning teams Axel has perfected the Dac-tag especially for the purification of recombinant proteins from insect cells. Using this methodology Axel and his team have been able to obtain incredibly pure preparations of enzymes from insect cells much better than could be achieved with other tags such as His, maltose binding and GST. We anticipate that the Dac-tag will become the frontline choice for the future purification of proteins from insect cells.
The Dac-tag is based on a fragment of penicillin binding protein 5, which binds in a pseudo reversible manner to ampicillin Sepharose. In order to obtain a pure protein of interest, the Dag-tag is cloned to the N or C-terminal end of the protein. Because of the properties of the Dac-tag, this fusion protein can then be readily isolated in a highly purified manner using ampicillin Sepharose, which is very selective for penicillin binding proteins. The Dac-tag has the advantage that it is monomeric and does not rely on Cysteine chemistry or metal affinity, so it is ideal for purification of proteins in the Ubiquitin arena.
The paper describing the development and use of the Dac Tag is now published: ‘The Dac-tag, an affinity tag based on penicillin binding protein 5.‘ in Analytical Biochemistry: Methods of the Life Sciences in press. Click here to access the paper.
The Dac-tag is based on a fragment of penicillin binding protein 5, which binds in a pseudo reversible manner to ampicillin Sepharose. In order to obtain a pure protein of interest, the Dag-tag is cloned to the N or C-terminal end of the protein. Because of the properties of the Dac-tag, this fusion protein can then be readily isolated in a highly purified manner using ampicillin Sepharose, which is very selective for penicillin binding proteins. The Dac-tag has the advantage that it is monomeric and does not rely on Cysteine chemistry or metal affinity, so it is ideal for purification of proteins in the Ubiquitin arena.
The paper describing the development and use of the Dac Tag is now published: ‘The Dac-tag, an affinity tag based on penicillin binding protein 5.‘ in Analytical Biochemistry: Methods of the Life Sciences in press. Click here to access the paper.