DES™-Inducible Kit with pCoBlast
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Citations & References (1)
Invitrogen™

DES™-Inducible Kit with pCoBlast

The DES™-Inducible Kit provides the expression vector pMT/V5-His and a choice of Blasticidin (pCoBlast) or hygromycin (pCoHygro) selection. This vectorRead more
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Catalog NumberQuantity
K5120011 kit
Catalog number K512001
Price (EUR)
2.745,00
Each
Add to cart
Quantity:
1 kit
Price (EUR)
2.745,00
Each
Add to cart
The DES™-Inducible Kit provides the expression vector pMT/V5-His and a choice of Blasticidin (pCoBlast) or hygromycin (pCoHygro) selection. This vector uses the Drosophila metallothionein gene promoter that is induced in S2 cells upon addition of copper sulfate or cadmium chloride to the culture medium. The pMT/V5-His vector offers the following features:

• Small size (3.5 kb) to improve DNA yields and increase subcloning efficiency
• C-terminal V5 epitope tag for rapid detection with Invitrogen's 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 coding sequence of the C-terminal tag.
For Research Use Only. Not for use in diagnostic procedures.
Specifications
Product TypeInducible 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
Each DES™-Inducible Kit contains 20 μg each of a selection vector (pCoHygro or pCoBlast), pMT/V5-His A, B, C, and pMT/V5-His/lacZ, Calcium Phosphate Transfection Kit, forward and reverse primers (2 μg each), frozen S2 cells, GIBCO™ Schneider's Drosophila Medium, 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.

Citations & References (1)

Citations & References
Abstract
A novel allelic variant of the human TSG-6 gene encoding an amino acid difference in the CUB module. Chromosomal localization, frequency analysis, modeling, and expression.
Authors: Nentwich Hilke A; Mustafa Zehra; Rugg Marilyn S; Marsden Brian D; Cordell Martin R; Mahoney David J; Jenkins Suzanne C; Dowling Barbara; Fries Erik; Milner Caroline M; Loughlin John; Day Anthony J;
Journal:J Biol Chem
PubMed ID:11854277
Tumor necrosis factor-stimulated gene-6 (TSG-6) encodes a 35-kDa protein, which is comprised of contiguous Link and CUB modules. TSG-6 protein has been detected in the articular joints of osteoarthritis (OA) patients, with little or no constitutive expression in normal adult tissues. It interacts with components of cartilage matrix (e.g. hyaluronan ... More