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Fluorescent protein promoter reporters: on-demand reporting

On-demand reporter vectors are the next generation of promoter reporters. With these reporter systems, you can compensate effectively for reporter background without compromising your assay’s signal intensity. Low background and a bright signal are no longer mutually exclusive.

On-demand reporter vectors are the next generation of promoter reporters. With these reporter systems, you can compensate effectively for reporter background without compromising your assay’s signal intensity. Low background and a bright signal are no longer mutually exclusive.

The challenges: overcoming high background & low signal intensity

Traditional promoter reporter assays generally struggle with the fact that most promoters are not very “tight”. As a result, your promoter of interest may drive reporter expression even without being activated—for example, during the time between transfection and the start of your experiment. These preexisting reporter molecules (the background) are the main cause of a low signal-to-noise ratio after promoter induction during the actual experiment.

One previous approach to this problem was to modify reporters for very quick, constitutive degradation. However, because these reporters are constitutively degraded as soon as they are made, it is impossible to accumulate a large quantity of reporter molecules inside the cell, even upon promoter activation. As a result, only a fraction of the reporter molecules are present long enough to be measured, and this type of assay has low signal intensity.

The solution: reporters on demand

The On-Demand Living Colors Fluorescent Protein Reporter Systems meet the challenge by providing both a low background and a broad dynamic range. This versatility is possible because they use a combination of technologies: each system includes a bright fluorescent protein reporter (AmCyan1, tdTomato, or ZsGreen1) for high signal intensity, coupled with ligand-dependent ProteoTuner protein stabilization/destabilization technology to eliminate background.

In these systems, the fluorescent protein reporter is expressed as a fusion protein tagged on its N-terminus with a ligand-dependent destabilization domain (DD). The DD rapidly targets the reporter protein for proteasomal degradation, guaranteeing a low reporter background signal at the start of your experiment. However, when the small, membrane-permeant ligand Shield1 is added to the sample, it binds to the DD and protects the reporter from degradation, so that it can accumulate.

By adding Shield1 simultaneously with your candidate inducer, you can effectively stabilize the reporter protein when it is synthesized in response to promoter activation. The majority of the fluorescent protein reporter molecules expressed during promoter activation will contribute to your readout, allowing for a considerably higher dynamic range and drastically improved signal-to-noise ratio compared to other types of reporter systems.

On-demand fluorescent protein reporter systems Traditional reporter systems
Background Uniformly low Promoter-dependent; may be high
Signal Bright Reporter-dependent. If background is low, signal is usually dim
Signal-to-noise ratio High, due to bright signal and low background Often low, especially when background is low
Studying timely promoter activity by eliminating unwanted reporter molecules Easy—simply remove Shield1 reagent Difficult; depends on reporter's natural lifespan

High signal, low background

In order to demonstrate the high signal-to-noise ratio and wide dynamic range of the DD fluorescent protein reporter systems, we compared the fold induction achieved using the DD-fluorescent protein reporters with that achieved using regular (non-destabilized) fluorescent proteins. The DD-tagged reporters stabilized by Shield1 had a much wider dynamic range, and therefore a much larger fold increase in the signal than the untagged versions of the same reporters. For the untagged versions, we observed high background fluorescence from reporter molecules that accumulated prior to induction, which drastically reduced the fold increase in signal intensity.

Choose your on-demand reporter system

Our on-demand reporter systems each consist of the necessary vectors (red, green, or cyan) plus Shield1.

Flexible choices

Use the DD fluorescent protein reporter systems to monitor any promoter of interest—just insert your promoter of interest upstream of the DD reporter. Choose from plasmid or lentiviral vector formats.

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Cat. # Product Size Price License Quantity Details
631089 CRE DD Cyan Reporter System Each USD $569.00

The CRE DD Cyan Reporter System is designed to monitor cAMP response element binding protein (CREB) activity in mammalian cells, with minimal background signal. It includes the pCRE-DD-AmCyan1 Reporter vector and Shield1.

pCRE-DD-AmCyan1 encodes a cyan fluorescent protein reporter tagged at its N-terminus with the ProteoTuner destabilization domain (DD), and under the control of the CRE promoter. The DD causes the DD-AmCyan1 reporter to be rapidly targeted to and degraded by proteasomes. This minimizes background fluorescence from leaky promoters prior to promoter activation.

To monitor CREB activity, a candidate inducer is added to the medium simultaneously with the DD's stabilizing ligand, Shield1. This allows DD-AmCyan1 to accumulate in response to CREB activation. As a result, only the reporter molecules expressed during CRE induction contribute to the fluorescence signal. This system provides a considerably higher signal-to-noise ratio than can be obtained with non-destabilized or constitutively destabilized reporter systems.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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631085 CRE DD Green Reporter System Each USD $569.00

The CRE DD Green Reporter System is designed to monitor cAMP response element binding protein (CREB) activity in mammalian cells, with minimal background signal. It includes the pCRE-DD-ZsGreen1 Reporter vector and Shield1.

pCRE-DD-ZsGreen1 encodes a green fluorescent protein reporter tagged at its N-terminus with the ProteoTuner destabilization domain (DD), and under the control of the CRE promoter. The DD causes the DD-ZsGreen1 reporter to be rapidly targeted to and degraded by proteasomes. This minimizes background fluorescence from leaky promoters prior to promoter activation.

To monitor CREB activity, a candidate inducer is added to the medium simultaneously with the DD's stabilizing ligand, Shield1. This allows DD-ZsGreen1 to accumulate in response to CREB activation. As a result, only the reporter molecules expressed during CRE induction contribute to the fluorescence signal. This system provides a considerably higher signal-to-noise ratio than can be obtained with non-destabilized or constitutively destabilized reporter systems.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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631087 CRE DD Red Reporter System Each USD $569.00 *

The CRE DD Red Reporter System is designed to monitor cAMP response element binding protein (CREB) activity in mammalian cells, with minimal background signal. It includes the pCRE-DD-tdTomato Reporter vector and Shield1.

pCRE-DD-tdTomato encodes a red fluorescent protein reporter tagged at its N-terminus with the ProteoTuner destabilization domain (DD), and under the control of the CRE promoter. The DD causes the DD-tdTomato reporter to be rapidly targeted to and degraded by proteasomes. This minimizes background fluorescence from leaky promoters prior to promoter activation.

To monitor CREB activity, a candidate inducer is added to the medium simultaneously with the DD's stabilizing ligand, Shield1. This allows DD-tdTomato to accumulate in response to CREB activation. As a result, only the reporter molecules expressed during CRE induction contribute to the fluorescence signal. This system provides a considerably higher signal-to-noise ratio than can be obtained with non-destabilized or constitutively destabilized reporter systems.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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632191 DD-AmCyan1 Reporter System Each USD $569.00

The DD-AmCyan1 Reporter System includes the pDD-AmCyan1 Reporter vector and Shield1. The pDD-AmCyan1 Reporter is a promoterless vector that allows you to insert your promoter of interest upstream of the cyan fluorescent protein AmCyan1, tagged at its N-terminus with the ProteoTuner destabilization domain (DD). The DD causes the reporter protein to be rapidly targeted to and degraded by proteasomes. This very efficient and controllable destabilization method will minimize any reporter background fluorescence from leaky promoters prior to promoter activation.

To analyze promoter activity, a candidate inducer is added to the medium along with Shield1, which effectively stabilizes the reporter protein and allows it to accumulate. As a result, only the reporter molecules expressed during promoter induction will contribute to the fluorescence signal, providing a considerably higher signal-to-noise ratio than that obtained with non-destabilized or constitutively destabilized reporter systems.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data
632190 DD-tdTomato Reporter System Each USD $569.00 *

The DD-tdTomato Reporter System includes the pDD-tdTomato Reporter vector and Shield1. The pDD-tdTomato Reporter is a promoterless vector that allows you to insert your promoter of interest upstream of the red fluorescent protein tdTomato, tagged at its N-terminus with the ProteoTuner destabilization domain (DD). The DD causes the reporter protein to be rapidly targeted to and degraded by proteasomes. This very efficient and controllable destabilization method will minimize background fluorescence from leaky promoters prior to promoter activation.

To analyze promoter activity, a candidate inducer is added to the medium along with Shield1, which effectively stabilizes the reporter protein and allows it to accumulate. As a result, only the reporter molecules expressed during promoter induction will contribute to the fluorescence signal, providing a considerably higher signal-to-noise ratio than that obtained with non-destabilized or constitutively destabilized reporter systems.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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632192 DD-ZsGreen1 Reporter System Each USD $569.00

The DD-ZsGreen1 Reporter System includes the pDD-ZsGreen1 Reporter vector and Shield1. The pDD-ZsGreen1 Reporter is a promoterless vector designed to insert your promoter of interest upstream of the green fluorescent protein ZsGreen1, tagged at its N-terminus with the ProteoTuner destabilization domain (DD). The DD causes the reporter protein to be rapidly targeted to and degraded by proteasomes. This very efficient and controllable destabilization method will minimize any reporter background fluorescence from leaky promoters prior to promoter activation.

To analyze promoter activity, a candidate inducer is added to the medium along with Shield1, which effectively stabilizes the reporter protein and allows it to accumulate. As a result, only the reporter molecules expressed during promoter induction will contribute to the fluorescence signal, providing a considerably higher signal-to-noise ratio than that obtained with non-destabilized or constitutively destabilized reporter systems.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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631748 Lenti-X™ DD Cyan Reporter System Each USD $1096.00 *

The Lenti-X DD Cyan Reporter System includes the Lenti-X DD-AmCyan1 Vector Set and Shield1. The Lenti-X DD-AmCyan1 Vector Set includes two lentiviral expression vectors (a reporter vector and a control vector) that produce high titers of recombinant lentivirus, which can efficiently transduce both dividing and nondividing mammalian cells. The system provides enough reagents for 16 packaging reactions. The pLVX-DD-AmCyan1 Reporter Vector is a promoterless vector that allows you to insert your promoter of interest upstream of the cyan fluorescent protein AmCyan1, tagged at its N-terminus with the ProteoTuner destabilization domain (DD). In the absence of the membrane-permeant ligand Shield1, the DD causes the reporter protein to be rapidly targeted to and degraded by proteasomes. This very efficient and controllable destabilization method minimizes reporter background fluorescence from leaky promoters prior to promoter activation.To analyze promoter activity, a candidate inducer is added to the medium along with Shield1, which binds to the DD tag and thereby stabilizes the reporter protein and allows it to accumulate. As a result, only the reporter molecules expressed during promoter induction will contribute to the fluorescence signal, providing a considerably higher signal-to-noise ratio than that obtained with nondestabilized or constitutively destabilized reporter systems. Cells used to monitor uninduced promoters (e.g., the negative control) will be treated with Shield1 alone.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data
631751 Lenti-X™ DD Green Reporter System Each USD $1096.00

The Lenti-X DD Green Reporter System includes the Lenti-X DD-ZsGreen1 Vector Set and Shield1. The Lenti-X DD-ZsGreen1 Vector Set includes two lentiviral expression vectors (a reporter vector and a control vector) that produce high titers of recombinant lentivirus, which can efficiently transduce both dividing and nondividing mammalian cells. The system provides enough reagents for 16 packaging reactions. The pLVX-DD-ZsGreen1 Reporter Vector is a promoterless vector that allows you to insert your promoter of interest upstream of the green fluorescent protein ZsGreen1, tagged at its N-terminus with the ProteoTuner destabilization domain (DD). In the absence of the membrane-permeant ligand Shield1, the DD causes the reporter protein to be rapidly targeted to and degraded by proteasomes. This very efficient and controllable destabilization method minimizes reporter background fluorescence from leaky promoters prior to promoter activation. To analyze promoter activity, a candidate inducer is added to the medium along with Shield1, which binds to the DD tag and thereby stabilizes the reporter protein and allows it to accumulate. As a result, only the reporter molecules expressed during promoter induction will contribute to the fluorescence signal, providing a considerably higher signal-to-noise ratio than that obtained with nondestabilized or constitutively destabilized reporter systems. Cells used to monitor uninduced promoters (e.g., the negative control) will be treated with Shield1 alone.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data
631753 Lenti-X™ DD Red Reporter System Each USD $1096.00 *

The Lenti-X DD Red Reporter System includes the Lenti-X DD-tdTomato Vector Set and Shield1. The Lenti-X DD-tdTomato Vector Set includes two lentiviral expression vectors (a reporter vector and a control vector) that produce high titers of recombinant lentivirus, which can efficiently transduce both dividing and nondividing mammalian cells. The system provides enough reagents for 16 packaging reactions. The pLVX-DD-tdTomato Reporter Vector is a promoterless vector that allows you to insert your promoter of interest upstream of the red fluorescent protein tdTomato, tagged at its N-terminus with the ProteoTuner destabilization domain (DD). In the absence of the membrane-permeant ligand Shield1, the DD causes the reporter protein to be rapidly targeted to and degraded by proteasomes. This very efficient and controllable destabilization method minimizes reporter background fluorescence from leaky promoters prior to promoter activation. To analyze promoter activity, a candidate inducer is added to the medium along with Shield1, which binds to the DD tag and thereby stabilizes the reporter protein and allows it to accumulate. As a result, only the reporter molecules expressed during promoter induction will contribute to the fluorescence signal, providing a considerably higher signal-to-noise ratio than that obtained with nondestabilized or constitutively destabilized reporter systems. Cells used to monitor uninduced promoters (e.g., the negative control) will be treated with Shield1 alone.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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631083 NFkappaB DD Cyan Reporter System Each USD $569.00

The NFkB DD Cyan Reporter System is designed to monitor NFkB activation in mammalian cells, with minimal background signal. It includes the pNFkB-DD-AmCyan1 Reporter vector and Shield1.

pNFkB-DD-AmCyan1 encodes a cyan fluorescent protein reporter tagged at its N-terminus with the ProteoTuner destabilization domain (DD), and under the control of the NFkB promoter. The DD causes the DD-AmCyan1 reporter to be rapidly targeted to and degraded by proteasomes. This minimizes background signal from leaky promoters prior to promoter activation.

To monitor NFkB activity, a candidate inducer is added to the medium simultaneously with the DD's stabilizing ligand, Shield1. This allows DD-AmCyan1 to accumulate in response to NFkB activation. As a result, only the reporter molecules expressed during NFkB induction contribute to the fluorescence signal. This system provides a considerably higher signal-to-noise ratio than can be obtained with non-destabilized or constitutively destabilized reporter systems.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data
631079 NFkappaB DD Green Reporter System Each USD $569.00

The NFkB DD Green Reporter System is designed to monitor NFkB activation in mammalian cells, with minimal background signal. It includes the pNFkB-DD-ZsGreen1 Reporter vector and Shield1.

pNFkB-DD-ZsGreen1 encodes a green fluorescent protein reporter tagged at its N-terminus with the ProteoTuner destabilization domain (DD), and under the control of the NFkB promoter. The DD causes the DD-ZsGreen1 reporter to be rapidly targeted to and degraded by proteasomes. This minimizes background signal from leaky promoters prior to promoter activation.

To monitor NFkB activity, a candidate inducer is added to the medium simultaneously with the DD's stabilizing ligand, Shield1. This allows DD-ZsGreen1 to accumulate in response to NFkB activation. As a result, only reporter molecules expressed during NFkB induction will contribute to the fluorescence signal. This system provides a considerably higher signal-to-noise ratio than can be obtained with non-destabilized or constitutively destabilized reporter systems.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data
631081 NFkappaB DD Red Reporter System Each USD $569.00 *

The NFkB DD Red Reporter System is designed to monitor NFkB activation in mammalian systems, with minimal background signal. It includes the pNFkB-DD-tdTomato Reporter vector and Shield1.

pNFkB-DD-tdTomato encodes a red fluorescent protein reporter tagged at its N-terminus with the ProteoTuner destabilization domain (DD), and under the control of the NFkB promoter. The DD causes the DD-tdTomato reporter to be rapidly targeted to and degraded by proteasomes. This minimizes background fluorescence from leaky promoters prior to promoter activation.

To monitor NFkB activity, a candidate inducer is added to the medium simultaneously with the DD's stabilizing ligand, Shield1. This allows DD-tdTomato to accumulate in response to NFkB activation. As a result, only the reporter molecules expressed during NFkB induction contribute to the fluorescence signal. This system provides a considerably higher signal-to-noise ratio than can be obtained with non-destabilized or constitutively destabilized reporter systems.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data

*You must be logged in to a Business Account in order to purchase these products online, since the purchase of these products may be restricted depending on your account type. Researchers at not-for-profit accounts receive a limited use license with their purchase of the product. Researchers at for-profit accounts must obtain a license prior to purchase. For details please contact licensing@takarabio.com.

Overview

  • The next generation of promoter reporters
  • High signal-to-noise ratio
  • Bright signal
  • Easy to monitor—red, green, and cyan options

More Information

Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.

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