DES™-Inducible/Secreted Kit with pCoBlast
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Invitrogen™

DES™-Inducible/Secreted Kit with pCoBlast

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)Read more
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Catalog NumberQuantity
K5130011 kit
Catalog number K513001
Price (EUR)
2.865,00
Each
Add to cart
Quantity:
1 kit
Price (EUR)
2.865,00
Each
Add to cart
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.
Specifications
Product TypeInducible/Secreted Kit
Protein Tag Position (to your gene)C-terminal
Quantity1 kit
VectorpMT
Cloning MethodGateway
Product LineDES
PromoterMT
Protein TagHis Tag (6x), V5 Epitope Tag
Unit SizeEach
Contents & Storage
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.

Frequently asked questions (FAQs)

What is the doubling time of Drosophila S2 cells?

When grown at the optimal temperature of 27-28ºC, Drosophila S2 cells grow very quickly, with an approximate 14-16 hr doubling time. The cells can also be grown at 22-25ºC with doubling time of 24hrs. Humidity is an issue, so a shallow pan of water can be left in the incubator, but the incubator does not need to be turned on.

Growth medium is also a factor in doubling time - the doubling times listed here are for Sf900 II medium.

The cells can reach maximum densities of 1 to 2.5 x 10E7 cells/ml in shake flask cultures.