Human NF-KappaB p65 Transcription Factor Activity Assay Kit

RayBiotech社のHuman NF-KappaB p65 Transcription Factor Activity Assay Kitです。
※本製品は研究用です。研究用以外には使用できません。

■Introduction

NF-κB is a transcription factor that plays a pivotal role in a number of physiological cell processes such as growth, apoptosis, and immune response through regulating the expression of a number of cellular genes. The NF-κB family includes five members: cRel, RelB, p65(RelA), p50(p105, precursor of p50), and p52(p100, precursor of p52), which form homo- or heterodimers when in active status. Normally, NF-κB is inactive and retained in the cytoplasm bound to its inhibitory protein IκB. Upon stimulation of various factors such as infection of pathogens including viruses and bacteria or treatment of cytokines such as IL-1 or TNF-a, IκB proteins are phosphorylated, ubiquitinated and finally degraded. Released NF-κB proteins then form homo- or heterodimers, most commonly p50/p65 or p50/p50, which are transported into the nucleus where they bind to specific DNA sequences of targeted genes and activate transcription. NF-κB also induces expression of its own inhibitor, IκBa, which binds to active NF-κB in the nucleus to terminate the induction of gene expression.

■Product Features

■Specificity

The olionucleotide/antibody pair provided in this kit recognizes human NF-κB p65 in whole lysates and nuclear extracts.

■Target Protein Information

Synonyms: Transcription factor p65 (Nuclear factor NF-kappa-B p65 subunit) (Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3)

Accession Number: Q04206

Gene Names: RELA NFKB3

Gene ID: 5970

■Species Detected

Human

■Application Notes

Kit Components

Other Materials Required

Protocol Outline

■Typical Data

Figure 1
Transcription factor activity assay of NF-κB p65 from nuclear extracts of HeLa cells or HeLa cells treated with TNF-a. After stimulation with TNF-a, activated NF-κB p65 is translocated into the nucleus where it binds with its corresponding DNA.

Figure 2
Transcription factor activity assay of NF-κB p65 from nuclear extracts of HeLa cells or HeLa cells treated with TNF-a with the specific competitor or non-specific competitor. The result shows specific binding of NF-κB p65 to the NF-κB DNA binding site.

Figure 3
Transcription factor activity assay of NF-κB p65 from nuclear extracts of HeLa cells treated with TNF-a and purified NF-κB p50. No cross-reactivity has been observed between NF-κB p65 and p50.

■Storage/Stability

Upon receipt, the positive control should be removed and stored at -20° or -80°C. The remainder of the kit can be stored for up to 6 months at 2-8°C from the date of shipment. Opened Microplate Wells or reagents may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack, reseal along entire edge.
Note: The kit can be used within one year if the whole kit is stored at -20°C upon receipt. Avoid repeated freeze-thaw cycles.

■ Antigen Information

Gene Symbol:

■ Product Specifications

Size: 1, 2, or 5 x 96-Well Strip Microplate Kit

■ Array/ELISA Format

Pathway:

Species Detected:

Solid Support: 96-well Microplate

Design Principle:

Quantitative/Semi-Quantitative: Semi-Quantitative

Method of Detection: Colorimetric

Compatible Sample Types: