pNL3.2.CMV Vector, 20 µg, Promega
Supplier: Promega Corporation
NanoLuc (Nluc) luciferase is a small bright luminescent reporter enzyme (19.1kDa) engineered for optimal performance. Use the pNL3.2.CMV Vector as a negative control in experiments measuring regulated changes in NanoLuc luciferase expression.
- Vector that Expresses NanoLuc(R) Luciferase Using the CMV Promoter
- Used as a control vector for the NanoLuc(R) fusion vectors
- Includes an optimized NanoLuc(R) luciferase gene that codes for a reporter protein with a shorter half-life (NlucP)
- Hygromycin resistance selectable marker means choosing transient transfection or generating a stable cell line
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. For use as a genetic reporter, multiple forms of NanoLuc luciferase have been configured to meet differing experimental objectives. NanoLuc-PEST (NlucP) present in pNL3.2.CMV Vector closely couples protein expression to changes in transcriptional activity and increased signal-to background ratios. As a result, the pNL3.2.CMV Vector is used as a negative control for experiments configured to measure regulated changes in NanoLuc luciferase expression levels. Luminescence is linearly proportional to the amount of NanoLuc protein over a 1,000,000-fold concentration range, with a signal half-life >/=2 hours when detected with Nano-Glo Luciferase Assay Reagent. NanoLuc luciferase possesses a number of physical properties that make it an excellent reporter protein: 1) very small, monomeric enzyme (171 amino acids; 513bp); 2) high thermal stability (Tm = 60°C); 3) active over a broad pH range (pH 6-8); 4) no post-translational modifications or disulfide bonds; 5) uniform distribution in cells; 6) emission spectrum well suited for bioluminescence resonance energy transfer (BRET; lambdamax = 465nM). NanoLuc luciferase is made available in a variety of plasmids designed for use in reporter gene assays of transcriptional control and with each of the NanoLuc forms (unfused Nluc, PEST destabilized NlucP, and secreted secNluc). The different pNL variations are designed for the following: 1) pNL1: cloning of a known or putative promoter region; 2) pNL2: cloning of a known or putative promoter region and establishment of a stable cell line through Hygromycin selection; 3) pNL3: cloning of a binding site or response element not in need of a basic promoter (such as are present in the pNL3.2.NF-kB-RE vector); 4) Control plasmids for the unfused, PEST-destabilized and secreted Nluc forms also are available. The pNL vector series uses a pGL4-based backbone for easy sequence transfer from existing plasmids. This backbone design also reduces anomalous results by removing many transcription factor binding sites and other potential regulatory elements. The NlucP gene is codon optimized and has removed many potential regulatory elements or other undesirable features (such as common restriction enzyme sites).
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