NF-kB p65 CLIA Kit

Catalog No. ABIN2866285

Product Details NF-kB p65 CLIA Kit

Target: NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))

Reactivity: Human

Detection Method: Chemiluminescent

Method Type: DNA-Binding ELISA

Application: ELISA

Purpose: DNA-binding ELISA that facilitate the study of transcription factor activation in mammalien tissue and cell culture extracts.

Brand: TransAM®

Sample Type: Cell Extracts, Tissue Samples

Analytical Method: Quantitative

Specificity: TransAM NFκB p65 Chemi Kits are tested for sensitivity in detecting NFκB activation.

Characteristics: Transcription factors are DNA-binding proteins that tightly regulate gene expression. They consist of two distinct domains - one that displays high affinity for a specific DNA sequence and one that confers transcriptional activity. Transcription factors are activated by phosphorylation of specific residues or by processing bound inhibitory proteins. Understanding and quantifying transcription factors is essential for the study of cell functions in relation to differentiation, brain activity, immune response, inflammation and various disease states. TransAM® Kits are sensitive, non-radioactive transcription factor ELISA kits that facilitate the study of transcription factor activation in mammalian tissue and cell extracts.
TransAM® Kits are DNA-binding ELISAs that facilitate the study of transcription factor activation in mammalian tissue and cell extracts. Each kit includes a 96-stripwell plate in which multiple copies of a specific double-stranded oligonucleotide have been immobilized. When nuclear or whole-cell extract is added, activated transcription factor of interest binds the oligonucleotide at its consensus binding site and is quantified using the included antibody, which is specific for the bound, active form of the transcription factor being studied.

Components: One or five 96-well plate(s) with plate sealer(s), primary antibody, HRP-conjugated secondary antibody, wild-type and mutated competitor oligonucleotides, positive control cell extract, DTT, Protease Inhibitor Cocktail, Lysis, Binding, 10X Washing and 10X Antibody Binding Buffers, and Developing and Stop Solutions.

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Comment:

Nuclear extracts prepared from Jurkat cells that are either unstimulated or stimulated with TPA + CI are diluted to 0.156 μg/well and assayed using the TransAM NFκB p65 Chemi Kit. The ratio of the signals from stimulated over unstimulated cells must be at least 4. Lot No. 26914050 gave a ratio of 22.8 (Figure 1). This ratio may vary depending on the basal level of NFκB activation in a given cell type. TransAM NFκB p65 Chemi Kits are also tested for specificity in detecting NFκB activation. TransAM NFκB assays are performed in the presence of an excess of oligonucleotide containing a wild-type or mutated NFκB consensus binding site (Figure 2). At 20X excess, the wild-type oligonucleotide prevents NFκB binding to the probe immobilized on the plate. Conversely, the mutated oligonucleotide has no effect on NFκB binding.

Assay Time: 5 h

Plate: Pre-coated

Restrictions: For Research Use only

Handling

Storage: 4 °C/-20 °C/-80 °C

Storage Comment: Except for the cell extract that must be kept at -80°C, the kit components can be stored at -20°C prior to first use. Then, we recommend storing the kit at 4°C except for the oligonucleotides, DTT, primary antibody, Herring sperm DNA and Protease Inhibitor Cocktail that should be kept at -20°C, and the cell extract at -80°C. This product is guaranteed for 6 months from date of receipt.

Expiry Date: 6 months

Target Details

Target: NF-kB p65 (NFkBP65) (Nuclear Factor-kB p65 (NFkBP65))

Alternative Name: Nfkb p65 (NFkBP65 Products)

Synonyms: C-Rel CLIA Kit, NFKB3 CLIA Kit, p65 CLIA Kit, NFkB CLIA Kit, c-rel CLIA Kit, zgc:100833 CLIA Kit, Xrel2 CLIA Kit, Xrel3 CLIA Kit, rel CLIA Kit, rel-A CLIA Kit, rel2 CLIA Kit, rel3 CLIA Kit, v-rel CLIA Kit, xrel CLIA Kit, REL proto-oncogene, NF-kB subunit CLIA Kit, RELA proto-oncogene, NF-kB subunit CLIA Kit, v-rel reticuloendotheliosis viral oncogene homolog A (avian) CLIA Kit, v-rel avian reticuloendotheliosis viral oncogene homolog CLIA Kit, v-rel avian reticuloendotheliosis viral oncogene homolog L homeolog CLIA Kit, REL CLIA Kit, RELA CLIA Kit, Rela CLIA Kit, Rel CLIA Kit, rel CLIA Kit, rel.L CLIA Kit

Pathways: NF-kappaB Signaling, RTK Signaling, TCR Signaling, TLR Signaling, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Cellular Response to Molecule of Bacterial Origin, Hepatitis C, Toll-Like Receptors Cascades, S100 Proteins