Cignal™ Lenti Positive Control (luc)

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Cignal™ Lenti Positive Control (luc)

Description

The Cignal Lenti Reporters are ready-to-transduce lentiviral particles for assessing cell signaling activities in virtually any mammalian cell type. The Cignal Lenti positive controls are ready-to-transduce lentiviral particles constitutively expressing firefly luciferase. This is an essential positive control for measuring transduction efficiency and optimizing reporter delivery. It can be used to troubleshoot problems with firefly luciferase assays in your cell system. The Cignal Lenti positive control (luc) is produced from the lentiviral backbone vector containing a gene encoding firefly luciferase driven by the CMV enhancer/promoter.

Why Cignal Lenti Positive Control (luc)?

Quantify transduction efficiency with luciferase activity
Optimize transduction conditions for your cell system
Help in troubleshooting luciferase assay in your cell system

For more information about the Cignal Lenti Reporters, including performance data, please visit the Cignal Lenti Reporter Assay home page.

Useful Links

Pricing and Ordering	Response Element Sequence	Other Lenti Reporter
User Manual	White Paper	Transfection Reagent
Register for Webinar	Technical Support

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Materials Included / Packing List:

Please check the kit components immediately after you receive this package. SABiosciences is only responsible for missing items reported within two (2) business days of receipt.

Kit Contents:

Component Specification Concentration* (total volume)

Positive Control firefly luciferase Reporter - 1 tube Ready-to-transduce lentivirus constitutively expressing firefly luciferase ≥ 0.8 X 10⁷ TU/ml (250 µl)**

Positive Control firefly luciferase Reporter - 8 tube Ready-to-transduce lentivirus constitutively expressing firefly luciferase ≥ 0.8 X 10⁷ TU/ml (2000 µl)***

* exact titer specified on Certificate of Analysis

** Supplied material provides sufficient lentivirus to transduce 2.5X10⁶ cells at a multiplicity of infection (MOI) of 1 TU per cell (refer to Cignal Lenti Reporter User Manual).

*** Supplied material provides sufficient lentivirus to transduce 2X10⁷ cells at a multiplicity of infection (MOI) of 1 TU per cell (refer to Cignal Lenti Reporter User Manual).

Storage Conditions: The Cignal Lenti Reporters are shipped on dry ice. Store all tubes at -80 ºC.

Biosafety Features

The Cignal Lenti Reporters are ready-to-transduce, replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles. The Cignal Lentiviral particles are safe to use. It is recommended that they be treated as Risk Group Level 2 (RGL-2) organisms. Follow all published RGL-2 guidelines for handling and waste decontamination. Details on the requirements for creating a BSL-2 work environment are available in the U.S. Department of Health and Human Services publication Biosafety in Microbiological and Biomedical Laboratories, 4th edition.

The biosafety features engineered into these vectors include:

A deletion in the promoter/enhancer region of the U3 portion of 3'LTR ensures self-inactivation of the lentiviral construct after transduction and integration into the genomic DNA of target cells.
The Cignal Lentiviral vector and plasmids expressing packaging proteins contain no significant areas of homology, thereby minimizing any chance for recombination.
None of the HIV-1 genes (gag, pol, rev) will be expressed in transduced cells, as they are expressed from packaging plasmids lacking packaging signal. Therefore, the lentiviral particles that are generated are replication-incompetent
No virulence genes ( vpr, vif, vpu and nef) are present in the Cignal Lentiviral vector.

Feature Function

RSV-5' LTR; Hybrid Rous sarcoma Virus (RSV) enhancer/promoter-U5 long terminal repeat Permits viral packaging and reverse transcription of viral mRNA

Psi; Packaging signal Allow viral packaging

RRE; Rev response element Involved in packaging of viral transcript

cppt; Central polypurine tract Involved in nuclear translocation and integration of transduced viral genome

luc (firefly luciferase) Allow quantification of transduction efficiency

hPGK; human phosphoglycerate kinase eukaryotic promoter Permits high-level expression of the mammalian selection marker (puromycin)

PuroR; puromycin resistance gene Can be used for mammalian selection

SIN/3'LTR; 3' self inactivating long terminal repeat Modified 3'LTR that allows viral packaging but self inactivates the 5'LTR for biosafety purpose. The element also contains a polyadenylation signal for efficient transcription termination

f1 ori; f1 origin of replication Allows episomal replication of plasmid in eukaryotic cells

AmpR; ampicillin resistance gene Allows selection of the plasmid in E.coli

CMV promoter Permits high level expression of reporter gene