Transfection Collection™ : NF-κB Transient Pack ( NF-κB Signaling Pathway)

Catalog #
79268
$780 *
Size: 100 reactions
Qty
*US Pricing only. For international pricing, please contact your local distributor.
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Description

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

The key to the NF-κB Transient Pack is the NF-κB luciferase reporter vector. This reporter contains a firefly luciferase gene under the control of multimerized NF-κB responsive element located upstream of a minimal promoter. The NF-κB reporter is premixed with constitutively-expressing Renilla luciferase vector, which 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 a firefly luciferase gene under the control of a minimal promoter, without any additional response elements. The negative control is critical to determining pathway specific effects and background luciferase activity.

Additionally, the pack includes cell culture medium (BPS Medium 1) that has been optimized for use with HEK293 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 TWO-Step Luciferase (Firefly & Renilla) Assay System. These reagents provide highly sensitive, stable detection of firefly luciferase activity and Renilla luciferase activity.  The TWO-step 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, but an alternate cell culture medium may be required for optimal cell growth.

This product has been cited 4 times.

Purchase of this cell line is for research purposes only; commercial use requires a separate license. View the full terms and conditions.

Synonyms
NF-kB reporter, NFkappaB reporter, luciferase reporter, NF-kB reporter, NFkB reporter, nfkb, nf-kb
Product Info
Storage and Usage
Citations4
Format

Concentration
Both the Reporter and Negative Control Reporter are provided at 60 ng DNA/µl (500 µl each)
Background

NF-κB (Nuclear Factor-KappaB) is a heterodimeric protein composed of different combinations of members of the Rel family of transcription factors. The NF-κB / Rel family of transcription factors (p50, p65, c-Rel, etc.) are involved in stress, immune, and inflammatory responses. In unstimulated cells, the NF-κB dimers are sequestered in the cytoplasm by inhibitory IκB proteins. Proinflammatory cytokines, LPS, growth factors, and antigen receptors activate IκB kinase (IKK), which phosphorylates the IκB proteins. Phosphorylation of IκB leads to its degradation, freeing NF-κB complexes to translocate to the nucleus, bind to NF-κB DNA response elements, and induce the transcription of the target genes.

References

1. Pessara, U., Koch, N. (1990). TNF alpha regulates expression of the major histocompatibility complex class II-associated invariant chain by binding of an NF-kappa B-like factor to a promoter element. Mol Cell Biol. 10(8):4146-4154.

2. Baeuerle, P.A. (1998). Pro-inflammatory signaling: last pieces in the NF-kappaB puzzle? Curr Biol. 8(1):R19-R22.

Application Reference(s):

1. Origin of Monocytes/Macrophages Contributing to Chronic Inflammation in Chagas Disease: SIRT1 Inhibition of FAK-NFκB-Dependent Proliferation and Proinflammatory Activation of Macrophages (2019)
2. PARP1-cGAS-NF-κB pathway of proinflammatory macrophage activation by extracellular vesicles released during Trypanosoma cruzi infection and Chagas disease (2020)
3. Origin of Monocytes/Macrophages Contributing to Chronic Inflammation in Chagas Disease: SIRT1 Inhibition of FAK-NFkB-Dependent Proliferation and Proinflammatory Activation of Macrophages. (2020)