|His tag vectors|
The polyhistidine tag is one of the most widely used tags. It can be provided on the N- or C-terminus of target proteins and can be used for protein purification by immobilized metal affinity chromatography and for immunodetection. Target proteins can be provided with a cleavable N-terminal 6xHis tag using our pAB-6xHis™ vector, or with a C-terminal FLAG-8xHis tag using pVL-FH vector.
The pAB-6xHis-MBP™ vector provides a cleavable N-terminal 6xHis-MBP tag. MBP (maltose binding protein) moiety is well documented to increase yield and solubility of target fusion proteins. Furthermore, it ensures exposure of the 6xHis tag on the surface of the fusion protein, which facilitates affinity purification under native conditions.
Another vector, pAB-bee™-8xHis is ideal for the expression of polyhistidine-tagged proteins which are translocated into the endoplasmic reticulum (ER), such as secreted and transmembrane proteins. Using this vector, the target protein can be provided with a strong signal sequence to ensure its efficient translocation into the ER, followed by a signal peptidase cleavage site and an N-terminal 8xHis tag. Optionally the 8xHis tag can be also provided on the C-terminus of a target protein. A similar pAB-bee vector encodes a strong signal sequence and an 8xHis tag which is provided on the C- terminus of the translocated proteins, but it does not encode for the N-terminal tag. A derivative of this vector pAB-bee™-FH encodes strong signal sequence and C-terminal dual FLAG-8xHis tag.
The plasmid transfer vectors described above can be combined with any of the listed below linearized baculovirus DNA vectors, i.e. ProEasy™, ProGreen™, ProFold™-ER1, ProFold™-C2, ProFold™-PDI, and ProFold™-0. Click on TECHNOLOGY for more information on baculovirus expression system. Go to BACULOVIRUS TUTORIAL for simple on-line instructions on baculovirus transfection, propagation of recombinant baculoviruses and recombinant baculovirus protein expression studies.
*The above plasmid transfer vectors are compatible with linearized baculovirus DNA from several other suppliers, e.g. BacMagic™, BacMagic™-2, BacMagic™-3, BacVector™-1000, BacVector™-2000, BacVector™-3000 (Novagen); BaculoGold™, BaculoGold™ bright (BD Pharmingen); flashBACGOLD™, flashBACULTRA™, flashBAC™ (OET); BacPAK6™ (Clontech); Sapphire™ (Orbigen). They are not compatible with Bac-to-Bac® and BaculoDirect™ DNA from Invitrogen. BacMagic™, BacMagic™-2, BacMagic™-3, BacVector™-1000, BacVector™-2000, BacVector™-3000 are trademarks of Novagen; BaculoGold™, BaculoGold™ bright are trademarks of BD Pharmingen; flashBACGOLD™, flashBACULTRA™, flashBAC™ are trademarks of OET; BacPAK6™ is a trademark of Clontech; Sapphire™ is a trademark of Orbigen, Bac-to-Bac® and BaculoDirect™ are trademarks of Invitrogen.
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|If you have not worked with recombinant baculoviruses, you can get a quick update at TECHNOLOGY. Go to BACULOVIRUS TUTORIAL for simple on-line instructions on baculovirus transfection, propagation of recombinant baculoviruses and recombinant baculovirus protein expression studies.|
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