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
Product	Product no.	Cat. no.	List price:
Cignal Finder Signal Transduction 45-pathway Reporter Array 200 ng of 10 or 45 dual-luciferase reporter assays in plate format, including positive and negative control assays show details	336821	CCA-901L	varies	configure

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.

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.