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Cignal Reporter Assay Kits

For rapid, sensitive, and quantitative assessment of signal transduction pathway activation

  • Rapid analysis of signal transduction pathway regulation
  • Exceptional sensitivity and specificity
  • Transfection-ready constructs
  • Includes positive and negative controls
  • Real-time live cell quantification (GFP format)

Cignal Reporter Assays provided in the Cignal Reporter Assay Kit enable rapid, sensitive, and quantitative assessment of signal transduction pathway activation by measuring the activities of downstream transcription factors. Two reporter systems are available: dual-luciferase format and GFP format.

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Product Product no. Cat. no. List price:
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-001L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-0020G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-0025L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-002G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-002L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-0033G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-0038L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-003L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-0041L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-004L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-005L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-006L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-007L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-008G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-008L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-009L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-010G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-010L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-011G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-011L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-012L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-013G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-013L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-014L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-015L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-016L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-017G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-017L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-018G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-018L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-1001G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-1014G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-1019L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-1022L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-1027G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-1035L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-2029L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-2032L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-2045L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-3021G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-3026L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-3039L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-3042L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-4023L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-4036L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-5020L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-5033L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-6022G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-6027L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-6030L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-6043L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-7024L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-7037L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-7040L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-7045G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-8021L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-8034L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-9028L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-9031L Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-9036G Inquire
For 500 reporter assays, 500 negative control assays, and 250 positive control assays; 500 µl (100 ng/µl) Reporter (inducible transcription-factor-responsive GFP reporter); 500 µl (100 ng/µl) Negative control (GFP reporter construct with GFP expression controlled by a minimal promoter); 250 µl (100 ng/µl) Positive control (constitutively expressing GFP construct)
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336841 CCS-9044L Inquire
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Cignal Reporter Assay Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


0
TNFα activates NFkB signaling activity in a dose-dependent manner.
293 H cells were transfected with Cignal NFkB Reporter Assay, negative or positive control. After 24 hours, cells were treated with different doses of hTNFα for a further 24 hours. Dual-luciferase assays were performed, and reporter activity values are expressed as arbitrary units using a Renilla reporter for internal normalization. Experiments were performed in triplicate and standard deviation is shown.
1
Assessing RNA interference phenotypes.

Cignal p53 Reporter Assays showed that p53 siRNA treatment abolished p53 transcription activity. HCT 116 cells were transfected with p53 reporter, negative control, and positive control, together with p53 siRNA or negative control siRNA. Dual-luciferase assays were performed. Reporter activity values are expressed as arbitrary units using a Renilla reporter for internal normalization. Experiments were performed in triplicate and the standard deviation is shown.

2
Assessing overexpression phenotypes.

Cignal RBP-Jκ Reporter Assay showed upregulation of Notch signaling activity after overexpression of activated Notch1. 293 H cells were transfected with RBP-Jκ reporter, negative control, and positive control. After 24 hours, cells were infected with 100 MOI of recombinant adenoviruses expressing activated Notch1 (Ad-NICD) or 100 MOI of recombinant adenovirus expressing GFP (Ad-GFP) for another 18 hours. Dual-luciferase assays were performed. Reporter activity values are expressed as arbitrary units using a Renilla reporter for internal normalization. Experiments were performed in triplicate and the standard deviation is shown.

3
Verification of small molecule drug candidate.

Cignal Retinoic Acid Response Element (RARE) Reporter Assay reported elevated retinoic acid receptor pathway activity after the treatment of all trans-retinoic acid (ATRA). CHO-K1 cells were transfected with RARE reporter, negative control, and positive control. After 16 hours, medium was changed to assay medium. After 24 hours, cells were treated with 1 µM all trans-rectinoic acid (ATRA) for 6 hours. Dual-luciferase assay was performed, and promoter activity values are expressed as arbitrary units using a Renilla reporter for internal normalization. Experiments were performed in triplicate and the standard deviation is shown.

4
Cignal Reporter Assay shows that hTNFa activated NFkB signaling pathway.
293 H cells were transfected with the Cignal NFkB-GFP Reporter Assay or negative control. After 16 hours, medium was changed to assay medium. After 24 hours, cells were treated with 10 ng/ml TNFα. After 18 hours, medium was removed from the wells and fluorescence activity was measured using a fluorometer. The fluorescence activity present in treated and nontreated negative control wells was subtracted from the fluorescence activity in treated and nontreated Cignal NFkB-GFP Reporter Assay wells and relative fluorescence activities expressed as arbitrary units. Experiments were performed in triplicate and the standard deviations are shown.
5
Cignal Reporter Assay measures activation of serum response factor (SRF) transcription activity.

293 H cells were transfected with the Cignal SRF-GFP Reporter Assay or negative control. After 16 hours, medium was changed to assay medium. After 24 hours, cells were treated with 10 ng/ml PMA and 10% serum. After 18 hours, medium was removed from the wells and fluorescence activity was measured using a fluorometer. The fluorescence activity present in treated and nontreated negative control wells was subtracted from the fluorescence activity in treated and nontreated Cignal SRF-GFP Reporter Assay wells and relative fluorescence activities expressed as arbitrary units. Experiments were performed in triplicate and the standard deviations are shown.

6
Cignal Reporter Assay procedure.
7
Cignal Reporter Assay principle.
Performance
Principle

Cignal Reporter Assays consist of multiple repeats of a specific transcription factor’s binding site and basic promoter elements to drive the expression of a reporter gene (firefly luciferase or GFP). These reporters are powerful tools in functional genomics and drug discovery for assessing pathway activity. When the pathway is activated or inhibited by a drug candidate, gene knockdown, overexpression, or peptide, luciferase or GFP reporter activity is modulated and can be measured quantitatively and rapidly (see figure "Cignal Reporter Assay principle"). The Cignal Reporter Assays are available as transfection-ready constructs utilizing two reporter systems: the dual-luciferase reporter or the GFP reporter (see flowchart "Cignal Reporter Assay procedure").

The dual-luciferase format is an end-point assay providing unsurpassed sensitivity, specificity, and signal-to-noise ratios. This format is available for all Cignal Reporter Assays. Each assay includes a pathway-focused transcription factor reporter and a noninducible negative control, as well as luciferase and GFP positive controls.

The GFP format is an outstanding method for monitoring live cell pathway regulation, with single cell resolution. It includes a pathway-focused transcription factor reporter and positive and negative controls.

Procedure

Cignal Reporter Assays include a preformulated, transfection-ready pathway reporter (dual-luciferase or GFP), plus a positive and negative control. The inducible pathway reporter and noninducible negative control are transfected and subjected to experimental treatments in parallel.

Dual-luciferase

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 transfectants serve as a specificity control. The positive control serves as a control for transfection efficiency, by monitoring firefly luciferase and Renilla expression.

GFP

GFP expression is quantified using a flow cytometer, fluorescence microscope, or fluorometer. The change in the activity of each signaling pathway is determined by comparing the GFP activities in treated versus untreated transfectants. The identically treated negative control transfectants serve as a specificity control. The positive control serves as a control for transfection efficiency, by monitoring GFP expression from the constitutively expressing CMV-GFP reporter.

Applications

Cignal Reporter Assays are powerful tools in functional genomics, proteomics, and drug discovery for assessing the biological impact of pathway inhibition or activation with siRNAs, proteins, and small molecule compounds.

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Brochures & Guides (1)
For cell-based analysis of pathway signaling activity
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Kit Handbooks (1)
Transfection Protocols (2)
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