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The pNLCoI Vectors are designed for use with the Nano-Glo Dual-Luciferase Reporter (NanoDLR) Assay System, which allows sequential detection of firefly and NanoLuc Luciferase in activity in the same sample. Both reagents provide stable glow-type luminescence signals with half-lives of approximately two hours allowing batch processing of samples and amenable to assays or screens in 96-, 384- or 1,536-well plate formats. Potent inhibition of firefly luciferase coupled with the high-intensity luminescence of NanoLuc luciferase maximizes sensitivity for detection of both reporters.

The phMGFP Vector contains the open reading frame for the Monster Green Fluorescent Protein cloned into a mammalian expression vector. The Monster Green Fluorescent Protein is encoded by an improved synthetic version of the green fluorescent protein gene originally cloned from Montastrea cavernosa. The synthetic gene (hMGFP) expresses a 26kDa protein that shows improved fluorescence intensity compared to the native gene. Furthermore, the hMGFP gene is codon optimized and cleared of most consensus sequence transcription factor binding sites to ensure reliability and high levels of expression. The Monster Green Fluorescent Protein generally fluoresces at least 20% brighter than other commercially available green fluorescent proteins (GFPs) and also reduces cytotoxicity, offering flexibility when working with transient and stable expression assays.

The pNL3.2.NF-kappaB-RE[NlucP/NF-kappaB-RE/Hygro] Vector contains five copies of an NF-kappaB response element (NF-kappaB-RE) that drives transcription of a destabilized form of NanoLuc luciferase, an engineered small (23.3kDa) luciferase fusion protein. The NlucP reporter consists of NanoLuc luciferase with a C-terminal fusion to PEST, a protein destabilization domain, which responds more quickly and with greater magnitude to changes in transcriptional activity than unmodified NanoLuc luciferase. NanoLuc (Nluc) luciferase is a small enzyme (19.1kDa) engineered for optimal performance as a luminescent reporter. The enzyme is about 100-fold brighter than either firefly (Photinus pyralis) or Renilla reniformis luciferase using a novel substrate, furimazine, to produce high intensity, glow-type luminescence. The luminescent reaction is ATP-independent and designed to suppress background luminescence for maximal assay sensitivity.

NanoLuc Stability Sensors are ready-to-use vector systems that enable stability studies of two key signaling proteins, HIF1A and NRF2, providing a method to directly measure this primary signaling event. The HIF1A Vector System enables simple quantification of intracellular HIF1A protein levels to study the dynamics of this signaling protein in mediating cellular response to hypoxia. It contains a vector encoding NanoLuc fused to the C-terminus of the HIF1A protein under control of the CMV promoter. The NRF2 Vector System enables simple quantification of intracellular NRF2 protein levels to study the dynamics of this signaling protein in mediating cellular response to oxidative stress. It contains a vector encoding NanoLuc fused to the C-terminus of the NRF2 protein under the control of the CMV promoter, a pKEAP1-expressing vector for proper regulation of intracellular NRF2 levels. Both systems include Transfection Carrier DNA for titratable intracellular fusion protein expression.

The pmirGLO Vector is designed to quantitatively evaluate microRNA (miRNA) activity by the insertion of miRNA target sites downstream or 3' of the firefly luciferase gene (luc2). Firefly luciferase is the primary reporter gene; reduced firefly luciferase expression indicates the binding of endogenous or introduced miRNAs to the cloned miRNA target sequence. This vector is based on Promega dual-luciferase technology, with luc2 used as the primary reporter to monitor mRNA regulation and Renilla luciferase (hRluc-neo) acting as a control reporter for normalization and selection.

The NanoLuc protein fusion vectors enable simple generation of N- or C-terminal fusions of NanoLuc luciferase with your protein of interest. The pNLF Vector series uses using traditional cloning with a multiple cloning site (MCS) to generate N- or C-terminal fusions to the full-length Nluc protein with the pNLF1-N or pNLF1-C [CMV/Hygro] Vectors or attach secreted Nluc to the N-terminus of the protein of interest with pNLF1-secN [CMV/Hygro] Vector. Also make NanoLuc protein fusions with the pF Vector series: Generate N- or C-terminal Nluc fusion proteins using the Flexi Vector Cloning System—a directional cloning method based on two rare-cutting restriction enzymes, SgfI and PmeI, that provides a rapid, efficient and high-fidelity way to transfer protein-coding regions between a variety of Flexi Vectors without the need to resequence.

The pRL Vectors are wildtype Renilla luciferase (Rluc) control reporter vectors that provide constitutive expression of Renilla luciferase and can be used in combination with a firefly luciferase vector to cotransfect mammalian cells. Expression of Renilla luciferase provides an internal control value to which expression of the experimental firefly luciferase reporter gene may be normalized. The pRL Vectors contain the cDNA encoding Renilla luciferase (Rluc) cloned from the anthozoan coelenterate Renilla reniformis (sea pansy). Four different promoter configurations are available. In general, we recommend validating the performance of specific co-reporter combinations in the desired target cells. In addition to the modified Rluc reporter gene, all pRL Vectors are isolated from a dam-/dcm- E. coli K host strain, allowing digestion with restriction enzymes that are sensitive to dam and dcm methylation.

The Promoter-driven firefly control vectors are commonly co-transfected with experimental Renilla or NanoLuc luciferase vectors for use in the Dual-Luciferase or Dual-Glo Reporter Assay Systems. The control firefly vectors should give an almost invariant level of activity, while the experimental Renilla or NanoLuc vector varies with treatment. The pGL4.13[luc2/SV40] ,pGL4.53[luc2/PGK] and pGL4.54[luc2/TK] Vectors offer three different promoter options for your dual-reporter assay.

The promoter-driven NanoLuc (Nluc) control vectors can be used to co-transfect with experimental firefly luciferase vectors when using the Nano-Glo Dual-Luciferase Reporter (NanoDLR) Assay System. NanoLuc luciferase is a small (19.1kDa), stable reporter enzyme that can be up to 100-fold more sensitive than the flash-type Renilla signal in the DLR Assay and more than 3,000-fold more sensitive than the Renilla signal in the Dual-Glo Assay. The increased brightness of the NanoLuc Luciferase means you use less control DNA, minimizing assay artifacts and providing a stable control signal for normalization of the experimental Fluc reporter. The vectors are engineered with minimal consensus transcription factor-binding sites to reduce anomalous expression.

The Promoter-driven Renilla control vectors are commonly co-transfected with experimental firefly luciferase vectors for use in the Dual-Luciferase or Dual-Glo Reporter Assay Systems. The control Renilla vectors should give an almost invariant level of activity, while the experimental firefly vector varies with treatment.

Promoterless firefly luciferase vectors are designed primarily to accept a putative promoter element for investigation of important regions controlling gene transcription. The promoterless vectors are available with three varieties of engineered firefly luciferase genes: luc2, luc2P or luc2CP. The luc2 gene is engineered to remove most cryptic transcription factor binding sites and improve mammalian expression through codon optimization. The luc2P, luc2CP and RapidResponse genes are luc2 genes appended with degradation sequences to influence the cellular half-life of the luc2 gene. The RapidResponse genes respond more rapidly to stimuli but at the expense of signal intensity. The luc2P (1-hour half-life) gene responds more rapidly than luc2 (3-hour half-life) with moderate signal intensity; the luc2CP (0.4-hour half-life) responds more quickly with the lowest signal intensity. The promoterless pGL4.10[luc2], pGL4.11[luc2P] and pGL4.12[luc2CP] Vectors do not have a selectable marker.

Promoterless firefly luciferase vectors are designed primarily to accept a putative promoter element for investigation of important regions controlling gene transcription. The promoterless vectors are available with three varieties of engineered firefly luciferase genes: luc2, luc2P or luc2CP. The luc2 gene is engineered to remove most cryptic transcription factor binding sites and improve mammalian expression through codon optimization. The luc2P, luc2CP and RapidResponse genes are luc2 genes appended with degradation sequences to influence the cellular half-life of the luc2 gene. The RapidResponse genes respond more rapidly to stimuli but at the expense of signal intensity. The luc2P (1-hour half-life) gene responds more rapidly than luc2 (3-hour half-life) with moderate signal intensity; the luc2CP (0.4-hour half-life) responds more quickly with the lowest signal intensity. The promoterless pGL4.14, pGL4.15 and pGL4.16 Vectors are available with the selectable marker hygromycin.

Promoterless firefly luciferase vectors are designed primarily to accept a putative promoter element for investigation of important regions controlling gene transcription. The promoterless vectors are available with three varieties of engineered firefly luciferase genes: luc2, luc2P or luc2CP. The luc2 gene is engineered to remove most cryptic transcription factor binding sites and improve mammalian expression through codon optimization. The luc2P, luc2CP and RapidResponse genes are luc2 genes appended with degradation sequences to influence the cellular half-life of the luc2 gene. The RapidResponse genes respond more rapidly to stimuli but at the expense of signal intensity. The luc2P (1-hour half-life) gene responds more rapidly than luc2 (3-hour half-life) with moderate signal intensity; the luc2CP (0.4-hour half-life) responds more quickly with the lowest signal intensity. The promoterless pGL4.17, pGL4.18 and pGL4.19 Vectors are available with a selectable marker, neomycin.

Promoterless firefly luciferase vectors are designed primarily to accept a putative promoter element for investigation of important regions controlling gene transcription. The promoterless vectors are available with three varieties of engineered firefly luciferase genes: luc2, luc2P or luc2CP. The luc2 gene is engineered to remove most cryptic transcription factor binding sites and improve mammalian expression through codon optimization. The luc2P, luc2CP and RapidResponse genes are luc2 genes appended with degradation sequences to influence the cellular half-life of the luc2 gene. The RapidResponse genes respond more rapidly to stimuli but at the expense of signal intensity. The luc2P (1-hour half-life) gene responds more rapidly than luc2 (3-hour half-life) with moderate signal intensity; the luc2CP (0.4-hour half-life) responds more quickly with the lowest signal intensity. The promoterless pGL4.20, pGL4.21 and pGL4.22 vectors are available with a selectable marker, puromycin.

The pNL1.1[Nluc] and pNL1.2[NlucP] Vectors are used to clone a putative promoter to drive expression of the bright NanoLuc reporter protein. NanoLuc (Nluc) luciferase is a small enzyme (19.1kDa) engineered for optimal performance as a luminescent reporter. The enzyme is about 100-fold brighter than either firefly (Photinus pyralis) or Renilla reniformis luciferase using a novel substrate, furimazine, to produce high intensity, glow-type luminescence. The luminescent reaction is ATP-independent and designed to suppress background luminescence for maximal assay sensitivity.

The pGL4.70[hRluc] Vector is designed primarily to accept a putative promoter element for investigation of important regions controlling gene transcription. Alternatively, this vector may be used as a promoterless control vector in a dual-reporter system with a firefly luciferase vector serving as the experimental vector. The promoterless vector is available with an hRluc engineered Renilla luciferase gene. The inserts cloned into this vector can easily be transferred using the multiple cloning site and a unique SfiI transfer scheme.

The pSV-Beta-Galactosidase Control Vector is a positive control vector for monitoring transfection efficiencies of mammalian cells. The SV40 early promoter and enhancer drive transcription of the lacZ gene, which encodes the beta-galactosidase enzyme. The pSV-Beta-Galactosidase Control Vector can be transfected individually or co-transfected with your DNA of interest. Beta-galactosidase is an excellent reporter enzyme that can be assayed quickly and directly in cell extracts using spectrophotometric, fluorescent or chemiluminescent assays. The pSV-Beta-Galactosidase Vector is a modification of pRSV-beta-Gal with SV40 and pUC18 sequences substituted for RSV and pBR322 sequences. The pSV-Beta-Galactosidase Vector will express beta-galactosidase in E. coli due to the presence of the E. coli gpt promoter located upstream of the lacZ gene. Colonies of E. coli containing the pSV-Beta-Galactosidase Vector will appear blue when plated on media containing X-Gal.