Pathway Focus: The NF-κB Pathway
- Fast detection and quantitation of transcription factors in cell lysates
- High-throughput assays in 96-well format
- Reproducible, quantitative results
Nuclear factor-kappa B (NF-κB) transcription factors are protein dimers composed of various combinations of members of the NF-κB/Rel protein family. Through its regulation of NF-κB, inhibitor of nuclear factor-κB α isoform (IκBα) controls immune and inflammatory responses, cell division, and apoptosis.
NF-κB mediates inflammatory responses, immune responses, responses to viral infections, cell division, and regulation of apoptosis, serving both as an anti-apoptotic signal and a pro-apoptotic signal.
ELISA technology allows a more detailed understanding of proteins in specific NF-кB pathways. We offer sandwich ELISA kits to quickly detect and quantify transcription factors in cell lysate models. These ELISA kits allow high throughput of samples in 96-well format and provide reproducible, quantitative results. Calibrated standard curves are provided to quantify the level of protein in each experimental run.
|Time course of TNF-α stimulation on Jurkat cells.
IκBα phosphorylation was observed 2 min after TNF-α treatment, followed by a rapid decrease due to subsequent degradation. The results correlate well with western blot analyses of the same samples (inset).
- Measure total and phosphorylated proteins
- Highly sensitive and specific antibodies
- Excellent lot-to-lot consistency with ABfinity™ technology
We offer highly specific ABfinity™ antibodies for NF-κB signal transduction research. These antibodies are validated against a variety of species for most applications.
- Find the Right Antibodies Using Our New Antibody Search Tool
Extracts of Jurkat cells were unstimulated (lane 1) or stimulated with 100 ng/mL PMA for 20 min, then with 0.5 μM Ca2+ ionophore for an additional 10 min (lanes 2–5) and resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was blocked with a 5% nonfat dried milk-TBST buffer for 1 hr at room temperature, then incubated with the NFκB [pS529] antibody for 2 hr at room temperature in a 3% nonfat dried milk-TBST buffer following prior incubation with no peptide (lanes 1, 2), the nonphosphopeptide corresponding to the phosphopeptide immunogen (lane 3), a generic phosphoserine-containing peptide (lane 4), or the phosphopeptide immunogen (lane 5). After washing, the membrane was incubated with goat F(ab′)2 anti-rabbit IgG HRP conjugate (Cat. No. ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that only the phosphopeptide corresponding to NFκB [pS529] blocks the antibody signal, demonstrating the specificity of the antibody. The data also show the induction of the phosphorylated signal after addition of PMA and a Ca2+ ionophore in this cell system.
Clonality, Clone (Isotype)
||pAb, ZK50 (Rb IgG)
||E, EMSA, WB
||mAb, 2A12A7 (Ms IgG2a)
||E, EMSA, WB
||mAb, 572 (Ms IgG1)
||Hu, Ms (B, Cn, Cp, Eq, Mk (Rh), Rt)
||IF, IP, WB
||pAb (Rb IgG)
||mAB 17.3 (Ms IgG1)
||IF, IP, WB
||mAb (Rb IgG)
||Hu (Ms, Sw)
E = ELISA; EMSA = electrophoretic mobility shift assay; IF = immunofluorescence; IP = immunoprecipitation; WB = western blot.
B = bovine; Cn = canine; Cp = chimpanzee; Eq = equine; Hu = human; Mk (Rh) = monkey (Rhesus); Ms = mouse; Rb = rabbit; Rt = rat; Sw = swine.