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Cignal™ Lenti Negative Control (GFP)

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 Negative controls (GFP) are ready-to-transduce lentiviral particles expressing GFP under the control of minimal CMV promoter. The negative control is critical to establishing the specificity of any treatment effects and determining background GFP fluorescence. Untreated cells transduced with Cignal Lenti Negative control (GFP) will provide a reference point for comparing all other samples. Treated cells transduced with Cignal Lenti Negative control (GFP) will provide a reference for specificity of response determined by Cignal Lenti reporters (GFP). The GFP (GFP) is an improved synthetic version of the green fluorescent protein gene. GFP has a excitation wavelength of 483nm and an emission wavelength of 502nm. GFP can be detected using common fluorescence filter sets or standard FACS settings as used for EGFP and FITC.

Why Cignal Lenti Negative Control (GFP)?

  • Establish the specificity of any treatment effects
  • Determine background GFP fluorescence
  • Test the sensitivity of target cells to lentiviral transduction

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 
Related Products
Cignal Lenti Reporter Assays shRNA plasmids Protein Phosphorylation ELISAs
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)
Negative Control GFP Reporter - 1 tube Ready-to-transduce lentivirus expressing GFP from a minimal promoter ≥ 0.8 X 107 TU/ml (250 µl)**
Negative Control GFP Reporter - 8 tube Ready-to-transduce lentivirus expressing GFP from a minimal promoter ≥ 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
GFP (Green Fluorescent Protein)  Allow visualization of background fluorescence
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 Origin of DNA replication for bacteriophage f1
AmpR; ampicillin resistance gene Allows selection of the plasmid in E.coli
TATA box Act as an minimal promoter