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Cignal 45-Pathway Reporter Array

For discovery of signaling pathway response to gene modification or chemical treatment

  • Analysis of multiple pathways in a single experiment
  • Transfection-ready protocol
  • Transfection and specificity controls included
  • Assays gene knockdown and overexpression
  • Assays chemical modulators

The Cignal 45-Pathway Reporter Array measures the activity of 45 signaling pathways. By screening pathway activities simultaneously, relevant pathways are quickly identified for further analysis. Cignal Finder Reporter Arrays pinpoint the pathways perturbed by a specific gene or drug. The plate format facilitates shorter transfection protocols and high-throughput analysis.

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Signal Transduction 45-Pathway Reporter Array
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200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays
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The Cignal 45-Pathway Reporter Array is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Cignal 45-Pathway Reporter Array procedure.
Principle

Each array includes 45 Cignal Reporter Assays and 2 controls in a 96-well plate format. Each reporter and control assay is dried down in each well of the plate.

Pathways and well positions in a Cignal 45-Pathway Reporter Array
Well position Pathway Transcription factor Well position Pathway  Transcription factor
 A1–2  Amino acid deprivation  ATF2/3/4  E1–2  MAP/JNK  AP-1
 A3–4  Androgen  AR  E3–4  MEF2  MEF2
 A5–6  Antioxidant response  Nrf1/Nrf2  E5–6  Myc  c-Myc
 A7–8  ATF6  ATF6  E7–8  Nanog  Nanog
 A9–10  C/EBP  C/EBP  E9–10  Notch  RBP-Jk
 A11–12  cAMP/PKA  CREB  E11–12  NFκB  NFκB
 B1–2  Cell cycle  E2F  F1–2  Oct4  Oct4
 B3–4  DNA damage  p53
 F3–4  Pax6  Pax6
 B5–6  EGR1  EGR1  F5–6  PI3K/Akt  FOXO
 B7–8  ER stress  CBF/NF-Y/YY1  F7–8  PKC/Ca2+  NFAT
 B9–10  Estrogen  ER  F9–10  PPAR  PPAR
 B11–12  GATA  GATA  F11–12  Progesterone  PR
 C1–2  Glucocorticoid  GR  G1–2  Retinoic acid  RAR
 C3–4  Heat shock  HSF-1  G3–4  Retinoid X  RXR
 C5–6  Heavy metal  MTF-1  G5–6  Sox2  Sox2
 C7–8  Hedgehog  Gli  G7–8  SP1  SP1
 C9–10  HNF4  HNF4  G9–10  STAT3  STAT3
 C11–12  Hypoxia  HIF-1α  G11–12  TGF-β  Smad2/3/4
 D1–2  Interferon regulation  IRF1  H1–2  Vitamin D  VDR
 D3–4  Type I interferon  STAT1/STAT2  H3–4  Wnt  TCF/LEF
 D5–6  Interferon gamma  STAT1  H5–6  Xenobiotic  AhR
 D7–8  KLF4  KLF4  H7–9  Negative control  
 D9–10  Liver X  LXR  H10–12  Positive control  
 D11–12  MAPK/Erk  SRF/Elk-1      


All reporter assays are based on dual-luciferase technology. Each reporter consists of a mixture of a pathway-focused transcription factor-responsive firefly luciferase construct and a constitutively expressing Renilla luciferase construct.

Dual-luciferase results are calculated for each transfectant. The change in the activity of each signaling pathway is determined by comparing the normalized luciferase activities of the reporter in treated versus untreated transfectants.

The identically treated negative control serves as a specificity control. The positive control serves as a control for transfection efficiency, by monitoring GFP expression, as well as a positive control for both the firefly and Renilla luciferase assays.

Procedure

The procedure comprises 3 simple steps (see figure "Cignal 45-Pathway Reporter Array procedure"):

Transfect Cignal Reporter Assays and test nucleic acids into cells 
Treat with siRNA, protein, peptide, or small molecule of interest 
Perform reporter quantitation using luciferase activity assays
Applications

The Cignal 45-Pathway Reporter Array is highly suited for research into phenotypes associated with RNAi or overexpression experiments, biological responses to small molecules or compounds, and mechanisms of action of proteins, peptides, and ligands.

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