Transfection Collection™ : AP1 Transient Pack (JNK Signaling Pathway)

Catalog #: 79266
Size: 500 reactions
$695
Background:

The stress-activated protein kinase/c-jun N-terminal kinase (SAPK/JNK) family of proteins includes mitogen-activated protein kinases (MAPKs) that are activated by stress, inflammatory cytokines, mitogens, oncogenes, and inducers of cell differentiation and morphogenesis. Upon activation of the SAPK/JNK pathway, MAP Kinase Kinases phosphorylate and activate JNKs. The activated JNKs translocate to the nucleus where they phosphorylate and activate transcription factors such as c-Jun. The activated c-Jun forms homodimers or heterodimers with fos family proteins which bind to the activator protein-1(AP1) response element and induce target gene transcription.

Description:

The AP1 Transient Pack is designed to provide the tools necessary for transiently transfecting and monitoring the activity of the JNK signaling pathway and the transcriptional activity of AP1 in cultured HEK293 cells. The kit contains transfection-ready vectors containing firefly luciferase as a JNK pathway-responsive reporter and constitutively expressing Renilla luciferase as a transfection control.  It also includes the Dual Luciferase detection reagents to detect both luciferase activities and specialized medium for growing and assaying HEK293 cells.

The key to the AP1 Transient Pack is the AP1 luciferase reporter vector. This reporter contains the firefly luciferase gene under the control of multimerized AP1 responsive elements located upstream of a minimal promoter. The AP1 reporter is premixed with a constitutively-expressing Renilla luciferase vector that serves as an internal control for transfection efficiency.

The pack also includes a non-inducible firefly luciferase vector premixed with constitutively- expressing Renilla luciferase vector as a negative control. The non-inducible luciferase vector contains the firefly luciferase gene under the control of a minimal promoter, without any additional response elements. The negative control is critical for determining pathway-specific effects and the background luciferase activity.

Additionally, the pack includes cell culture medium (BPS Medium 1) that has been optimized for use with HEK293 and HeLa cells*. BPS Medium 1 includes MEM medium, 10% fetal bovine serum, 1% non-essential amino acids, sodium pyruvate, and 1% Pen/Strep.  Finally, the pack provides the Dual Luciferase (Firefly-Renilla) Assay System. These luciferase reagents provide highly sensitive, stable detection of firefly luciferase activity and Renilla luciferase activity.  The dual luciferase reagents can be used directly in cells in growth medium, and can be detected with any luminometer; automated injectors are not required.          

*Note:  the kit may be used with other cell lines than HEK293 or HeLa, but an alternate cell culture medium may be required for optimal cell growth,

Synonym(s):
JNK reporter, SAPK reporter
Concentration:
Both the Reporter and Negative Control Reporter are provided at 60 ng DNA/µl (500 µl each)
Format:

Instructions for use:
See data sheet insert for detailed transfection and assay protocols.
Storage / Stability:
Stable at least 12 months from date of receipt, when stored as directed (-20°C)
Application(s):
• Monitor JNK signaling pathway activity and AP1-mediated activity.

• Screen for activators or inhibitors of the JNK signaling pathway.

• Study effects of RNAi or gene overexpression on the activity of the JNK pathway.

Notes:
Note: These vectors are ready-to-use for transient transfection. They are NOT SUITABLE for transformation and amplification in bacteria.
Warning(s):
Avoid freeze/thaw cycles
Product Type:
Assay Kit
Data shown is lot-specific. Contact us for specific information on other lots.

Quality Assurance

Product Datasheets

79266.pdf