NF-kB p65 ELISA Kit

Catalog No. ABIN2866268

Product Details NF-kB p65 ELISA Kit

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

Reactivity: Human

Detection Method: Colorimetric

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 Flexi NFκB p65 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 0.625 μg/well and assayed using the TransAM Flexi NFκB p65 Kit. The ratio of the signals from the stimulated over unstimulated cells must be above 2. Lot No. 11514049 was developed for 11 minutes. It gave a ratio of 2.7 (Figure 1). This ratio may vary depending on the basal level of NFκB activation in a given cell type. TransAM Flexi NFκB p65 Kits are also tested for specificity in detecting NFκB activation. TransAM Flexi NFκB assays are performed in the presence of an excess of competitor oligonucleotide containing a wild-type or mutated NFκB consensus binding site (Figure 2). At 10X excess, the wild-type oligonucleotide prevents NFκB binding to the biotinylated probe. 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, and Protease Inhibitor Cocktail that should be kept at -20°C. This product is guaranteed for 6 months from date of receipt.

Expiry Date: 6 months

Target Details for NF-kB p65

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

Alternative Name: Nfkb p65 (NFkBP65 Products)

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