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pCMV-GLuc Control Plasmid |
 | | | | pCMV-GLuc Control Plasmid | | | |
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Prices are in US dollars and valid only for US orders.
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- Naturally secreted — cell lysis not necessary
- Sensitivity — 1,000-fold higher than other luciferases
- Stability — can be stored for several days at 4°C without loss of activity
Description:
The pCMV-GLuc Control Plasmid contains the "humanized" coding sequence for the secreted Gaussia Luciferase (GLuc) under the control of the Cytomegalovirus (CMV) promoter, for constitutive expression in mammalian cells. pCMV-GLuc can be used as a positive control in transfection experiments alone or in combination with other reporter vectors.
Gaussia Luciferase is a new reporter luciferase originally isolated from the marine organism Gaussia princeps (1, 2). This luciferase, does not require ATP, and catalyzes the oxidation of the substrate coelenterazine in a reaction that emits light (470 nm).*
* The GLuc activity can be measured using the Gaussia Luciferase Assay Kit (NEB #E3300)
Figure: Restriction map of pCMV-GLuc Vector. Only unique restriction sites are shown.
Figure: Comparison of light activity obtained from secreted GLuc versus secreted Renilla Luciferases in U2OS cells transfected with the indicated amounts of the corresponding plasmids and TransPass™ D2 Transfection Reagent. 20 µl of cell culture supernatant from each well was measured with the Gaussia Luciferase Assay Kit in a Molecular Devices Luminometer.
Source:
Isolated from E. coli strain ER2272 by a standard DNA purification procedure.
Advantages:- GLuc possesses a natural secretory signal and upon expression is secreted into the cell medium of cultured mammalian cells. Therefore, lysis of the cells is not necessary before assaying activity. This allows for multiple samples to be assayed from the same transfected cells, as well as collecting data at different time points.
- GLuc generates over 1000-fold higher bioluminescent signal intensity than Firefly and Renilla Luciferases (2). GLuc is a very sensitive transcriptional reporter, ideal with assays that use low numbers of cells (e.g. 384 well plates) or difficult to transfect cells.
- The secreted protein is very stable and can be stored for several days at 4°C without loss of activity.
Concentration:
500 μg/ml
Storage Conditions
Storage Conditions:
10 mM Tris-HCl
1 mM EDTA
pH 7.5 @ 25°C
Storage Temperature:
-20°C
References
- Verhaegen, M. and Christopoulos, T.K. (2002) Anal. Chem., 74, 4378-4385.
- Tannous, B.A., Kim, D.E., Fernandez, J.L., Weissleder, R., and Breakefield, X.O. (2005) Mol. Ther., 11, 435-443.
Companion Products
Gaussia Luciferase Assay Kit
Luciferase Cell Lysis Buffer
pGLuc-Basic Vector
pGLuc-Reverse Sequencing Primer (24-mer)
pNEBR-X1GLuc Control Plasmid
TransPass™ COS/293 Transfection Reagent
TransPass™ D1 Transfection Reagent
TransPass™ D2 Transfection Reagent
TransPass™ HeLa Transfection Reagent
TransPass™ HUVEC Transfection Reagent
TransPass™ V
Legal
Licenses/Patents/Disclaimers:
The CMV promoter is covered under U.S. Patents 5,168,062 and 5,385,839 and its use is permitted for research purposes only. Any other use of the CMV promoter requires a license from the University of Iowa Research Foundation, 214 Technology Innovation Center, Iowa City, IA 52242.
By opening this package or by using the materials enclosed within:
Recipient is legally bound and accepts the following terms and conditions:
Non-Commercial Entities: This product is covered by US Patent 6,232,107 and other patents that are legal property as assigned to Prolume Ltd./Nanolight Technologies. This product is licensed only to the purchasing laboratory-research group. Recipient agrees not to transfer this plasmid or derivatives of this vector to any other laboratory, person or research group, even if within the same institution.
Recipient agrees not to alter or make any changes to the nucleotide coding sequence or secretory coding sequence of the luciferase(s) contained within without prior written permission from Prolume Ltd./Nanolight Technologies (www.nanolight.com). Recipient agrees not to file for any patent rights or to any inventions claiming any portion of the luciferase(s) within this material without prior written permission from Prolume Ltd./Nanolight Technologies.
Commercial For-Profit Entities & Non-Profit Foundations (herein referred to as Commercial Recipients): Commercial Recipients wishing to derive products, engage in the sale or license of any products, discover drugs or make inventions by use of the materials enclosed, fully agree to the terms mentioned above for Non-Commercial Entities; AND ADDITIONALLY agree to and are hereby bound to use the materials FOR EVALUATION PURPOSES ONLY. Commercial Recipient hereby agrees to destroy and cease use of any materials or derivatives containing any portion of these materials within 120 days from receipt. Commercial Recipient agrees not to use the materials for any use, other than the 120 day suitability evaluation without prior written permission or obtaining a valid license from Prolume Ltd. Nanolight Technologies.
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