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Pathway Focus: The NF-κB Pathway

Measure IкBα [pS32] and Total Protein
Measure IкBα [pS32] and Total Protein—phosphoELISA™ Assays
Antibodies for NF-kB Signaling PathwaysAntibodies for NF-κB Signaling Pathways—ABfinity™ Technology

 

Cell Staining Simulation tool

Primary Antibody Search Tool

3D Cell and Cell Staining Simulation tool iPad app

Measure IкBα [pS32] and Total Protein—phosphoELISA™ Assays

  • 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.

Disease states in which NF-κB is improperly regulated include cancer, neurodegenerative diseases, and numerous diseases related to immune system dysregulation such as arthritis, asthma, and inflammatory bowel disease.

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
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).
For research use only. Not intended for any animal or human therapeutic or diagnostic use, unless otherwise stated.
phosphoELISA™ Assays Selection Guide

Product Species
Quantity
Cat. No.
IκBα (total)
Human
96 tests
KHO0211
IκBα [pS32]
Human96 tests
KHO0221

Antibodies for NF-κB Signaling Pathways—ABfinity™ Technology

  • 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.
We also have more than 300 phosphorylation site–specific antibodies available that have been validated with peptide competition experiments. This offering includes antibodies that recognize NF-κB, RelB, TRAF6, IκBα, or related pathways such as Akt, TLC, and FAK.

Standard curve of Human SAA ELISA Kit
Peptide competition.

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.
ABfinity™ Recombinant Rabbit Monoclonal Antibody Selection Guide

Target Protein Clonality, Clone (Isotype)
Reactive Species Applications
Quantity
Cat. No.
NFκB (p50)
pAb, ZK50 (Rb IgG)
Hu
E, EMSA, WB
100 µg
513500
NFκB (p65)
mAb, 2A12A7 (Ms IgG2a)
Hu
E, EMSA, WB
100 µg339900
NFκB (p65)
mAb, 572 (Ms IgG1)
Hu, Ms (B, Cn, Cp, Eq, Mk (Rh), Rt)
IF, IP, WB
100 µg436700
NFκB [pS529]
pAb (Rb IgG)
Hu
WB
10 blots
44711G
RelB
mAB 17.3 (Ms IgG1)
Hu
IF, IP, WB
100 µg437500
IκB [pS32]
mAb (Rb IgG)
Hu (Ms, Sw)
E, WB
100 µL44725G

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.
For research use only. Not intended for any animal or human therapeutic or diagnostic use, unless otherwise stated.