The DES®-Inducible/Secreted Kit provides the vector pMT/BiP/V5-His for inducible, secreted expression of recombinant proteins and a choice of Blasticidin (pCoBlast) or hygromycin (pCoHygro) selection. This vector offers the inducible metallothionein promoter that is induced upon addition of copper sulfate or cadmium chloride. The N-terminal signal sequence from the insect BiP gene is provided to direct the recombinant fusion protein through the secretory pathway of S2 cells into the culture medium. The pMT/BiP/V5-His vector offers the following additional features:
• Small size (3.6 kb) to improve DNA yields and increase subcloning efficiency • C-terminal V5 epitope tag for rapid detection with Invitrogens Anti-V5 Antibody • C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin
To facilitate cloning, a set of three vectors-A, B, and C-is provided. Each vector has the multiple cloning site in a different reading frame relative to the BiP signal sequence.
For Research Use Only. Not for use in diagnostic procedures.
The DES® Inducible/Secreted Kits contains 20 µg of each of a selection vector (pCoHygro or pCoBlast), pMT/BiP/V5-His A, B, C and pMT/BiP/V5-His/GFP, Calcium Phosphate Transfection Kit, forward and reverse primers (2 µg each), GIBCO™ Schneider's Drosophila Medium, frozen S2 cells, and the appropriate selection agent. Store S2 cells in liquid nitrogen. Store medium and selection agent at +4°C. Store all other reagents at -20°C. All components are guaranteed stable for 6 months when properly stored.