Invitrogen™

pGlow TOPO™ TA Expression Kit

The pGlow TOPO® TA Expression Kit uses the Cycle 3 GFP mutant (1) as a reporter protein for transcriptional activity.Read more

Catalog Number	Quantity
K483001	20 reactions

Catalog number K483001

Price (EUR)

1.278,00

Quantity:

20 reactions

Price (EUR)

1.278,00

The pGlow TOPO® TA Expression Kit uses the Cycle 3 GFP mutant (1) as a reporter protein for transcriptional activity. Cycle 3 GFP is ideal for non-invasive analysis of promoter elements within intact, living cells. The fluorescent property of Cycle 3 GFP allows in vivo detection in virtually any cell type or species using microscopy with wild-type GFP filter sets or by fluorescence-activated cell sorting methods.

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

Specifications

Delivery TypeTransfection

For Use With (Application)Reporter Assays, Clone your own Promoter

Product TypeTOPO TA Expression Kit

Quantity20 reactions

Reporter GeneGFP (Cycle 3)

Selection Agent (Eukaryotic)Geneticin™ (G-418)

VectorpGlow, TOPO-TA Vectors

Cloning MethodTOPO-TA

Product LineTOPO

PromoterNone (Promoterless)

Protein TagUntagged

Unit SizeEach

Contents & Storage

pGlow TOPO™ TA Expression Kits contains two boxes. The TOPO™ TA Cloning box contains linearized and topoisomerase I-activated pGlow-TOPO™ vector, dNTPs, primers for sequencing, salt solution, and control PCR template and primers; store at -20°C. The One Shot™ box contains One Shot™ TOP10 E. coli cells, S.O.C. medium, and a supercoiled control plasmid; store at -80°C. The kits are guaranteed stable for 6 months when properly stored.

Frequently asked questions (FAQs)

Can I store my competent E. coli in liquid nitrogen?

We do not recommend storing competent E. coli strains in liquid nitrogen as the extreme temperature can be harmful to the cells. Also, the plastic storage vials are not intended to withstand the extreme temperature and may crack or break.

How should I store my competent E. coli?

We recommend storing our competent E. coli strains at -80°C. Storage at warmer temperatures, even for a brief period of time, will significantly decrease transformation efficiency.

Can I use pBlue- or pGlow-TOPO reporter vectors to evaluate promoter function in E. coli?

pBlue-TOPO contains a cryptic prokaryotic promoter upstream of the lacZ reporter gene, due to which E. coli transformants may appear to be light blue when screened on plates containing X-Gal. Hence, we do not recommend using pBlue-TOPO to evaluate promoter function in E. coli. However, pGlow-TOPO can be used for these studies. Note that background expression of beta-galactosidase from pBlue-TOPO does not occur in mammalian cells.

Do you offer a vector that I could use to do some promoter analysis studies?

Yes, we do offer the pBlue-TOPO and pGlow-TOPO vectors that facilitate cloning of the DNA sequence of interest directly upstream of either the b-galactosidase or Cycle 3 GFP gene, respectively.
pBlue-TOPO is ideal for functional analysis of promoters with low transcriptional activity, since assays for beta-galactosidase are easy to perform and are quantitative at very low levels of expression. pGlow-TOPO is ideal for non-invasive analysis of promoter elements within intact, living cells. The fluorescent property of Cycle 3 GFP allows in vivo detection in virtually any cell type or species using microscopy with wild-type GFP filter sets or by fluorescence-activated cell sorting methods.

Can GFP fluorescence be detected in cells that have been stained for beta-galactosidase?

We recommend looking for GFP fluorescence before staining for beta-galactosidase. This is because the beta-galactosidase staining process produces a very high autofluorescence that will interfere with detection of GFP fluorescence.

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