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Invitrogen™ DES™-Inducible/Secreted Kit with pCoBlast

Catalog No. K513001
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Catalog No. K513001 Supplier Invitrogen™ Supplier No. K513001
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Description

Includes

The Inducible/Secreted Kits contains 20μg 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 (2μg each), GIBCO Schneider's Drosophila Medium, frozen S2 cells, and the appropriate selection agent.

Provides vector pMT/BiP/V5-His for inducible, secreted expression of recombinant proteins and Blasticidin (pCoBlast) selection

  • This vector offers inducible metallothionein promoter that is induced upon addition of copper sulfate or cadmium chloride
  • N-terminal signal sequence from insect BiP gene is provided to direct recombinant fusion protein through secretory pathway of S2 cells into culture medium
  • pMT/BiP/V5-His vector offers small size (3.6kb) to improve DNA yields and increase subcloning efficiency
  • C-terminal V5 epitope tag for rapid detection withAnti-V5 Antibody
  • C-terminal 6xHis tag for simple purification of recombinant fusion proteins using nickel-chelating resin
  • To facilitate cloning, set of three vectors-A, B, and C-is provided: Each vector has multiple cloning site in different reading frame relative to BiP signal sequence

Inducible Protein Expression in Insect Cells, Insect Expression, Protein Expression, Proteins, Expression, Isolation and Analysis, Secreted Protein Expression in Insect Cells

Specifications

Promoter MT
Product Type Inducible/Secreted Kit
Content And 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.
Protein Tag Position (to your gene) C-terminal
Protein Tag His Tag (6x), V5 Epitope Tag
Cloning Method Gateway
Quantity 1 kit
Vector pMT
Product Line DES

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.

For Research Use Only. Not for use in diagnostic procedures.