Mouse IkappaB alpha ELISA Kit (ab206308)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 14.7 pg/ml
- Range: 46.875 pg/ml - 3000 pg/ml
- Sample type: Cell culture extracts, Tissue Extracts
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Mouse
Product nameMouse IkappaB alpha ELISA Kit
See all IKB alpha kits
Intra-assay Sample n Mean SD CV% cell extract 5 6.5% Inter-assay Sample n Mean SD CV% cell extract 3 10%
Sample typeCell culture extracts, Tissue Extracts
Assay typeSandwich (quantitative)
Range46.875 pg/ml - 3000 pg/ml
Sample specific recovery Sample type Average % Range Cell culture media 90.9 89.7% - 92.7%
Assay time1h 30m
Assay durationOne step assay
Species reactivityReacts with: Mouse
Mouse IkappaB alpha ELISA Kit (ab206308) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of IkappaB alpha protein in cell culture extracts and tissue extracts. It uses our proprietary SimpleStep ELISA® technology. Quantitate Mouse IkappaB alpha with 14.7 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
- Single-wash protocol reduces assay time to 90 minutes or less
- High sensitivity, specificity and reproducibility from superior antibodies
- Fully validated in biological samples
- 96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpleStep ELISA® kits.
IkappaB alpha inhibits the activity of dimeric NF-kappa-B/REL complexes by trapping REL dimers in the cytoplasm through masking of their nuclear localization signals. On cellular stimulation by immune and proinflammatory responses, IkappaB alpha becomes phosphorylated promoting ubiquitination and degradation, enabling the dimeric RELA to translocate to the nucleus and activate transcription.
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PlatformPre-coated microplate (12 x 8 well strips)
Storage instructionsStore at +4°C. Please refer to protocols.
Components 1 x 96 tests 10X Mouse IkappaB alpha Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml 50X Cell Extraction Enhancer Solution (ab193971) 1 x 1ml 5X Cell Extraction Buffer PTR (ab193970) 1 x 10ml Antibody Diluent 4BR 1 x 6ml Mouse IkappaB alpha Capture Antibody (lyophilized) 1 vial Mouse IkappaB alpha Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml
FunctionInhibits the activity of dimeric NF-kappa-B/REL complexes by trapping REL dimers in the cytoplasm through masking of their nuclear localization signals. On cellular stimulation by immune and proinflammatory responses, becomes phosphorylated promoting ubiquitination and degradation, enabling the dimeric RELA to translocate to the nucleus and activate transcription.
Involvement in diseaseEctodermal dysplasia, anhidrotic, with T-cell immunodeficiency autosomal dominant
Sequence similaritiesBelongs to the NF-kappa-B inhibitor family.
Contains 5 ANK repeats.
modificationsPhosphorylated; disables inhibition of NF-kappa-B DNA-binding activity. Phosphorylation at positions 32 and 36 is prerequisite to recognition by UBE2D3 leading to polyubiquitination and subsequent degradation.
Sumoylated; sumoylation requires the presence of the nuclear import signal. Sumoylation blocks ubiquitination and proteasome-mediated degradation of the protein thereby increasing the protein stability.
Monoubiquitinated at Lys-21 and/or Lys-22 by UBE2D3. Ubiquitin chain elongation is then performed by CDC34 in cooperation with the SCF(FBXW11) E3 ligase complex, building ubiquitin chains from the UBE2D3-primed NFKBIA-linked ubiquitin. The resulting polyubiquitination leads to protein degradation. Also ubiquitinated by SCF(BTRC) following stimulus-dependent phosphorylation at Ser-32 and Ser-36.
Deubiquitinated by porcine reproductive and respiratory syndrome virus Nsp2 protein, which thereby interferes with NFKBIA degradation and impairs subsequent NF-kappa-B activation.
Cellular localizationCytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm by a nuclear localization signal (NLS) and a CRM1-dependent nuclear export.
- Information by UniProt
- I kappa B alpha
- Entrez Gene: 18035 Mouse
- SwissProt: Q9Z1E3 Mouse
- Unigene: 170515 Mouse
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
Background-subtracted data values (mean +/- SD) are graphed.
The concentrations of IkappaB alpha were measured in duplicates, interpolated from the IkappaB alpha standard curves and corrected for sample dilution. Note that 1X Diluted A20 cell extract samples were at 50 µg/mL. The interpolated, dilution factor-corrected values are plotted in pg of IkappaB alpha per µg of total protein (mean +/- SD, n=2).
Cell extracts were prepared from untreated mouse A20, rat H9C2 and rat H4II2 cells, or mouse NIH/3T3 cells treated for 20 minutes with 20 ng/mL TNF-alpha or mock-treated. The concentrations of IkappaB alpha were measured in three dilutions in duplicates, interpolated from the IkappaB alpha standard curves and corrected for sample dilution. Note that 1X Diluted A20 cell extract samples were at 50 µg/mL. Note that 1X Diluted NIH/3T3 cell extract samples were at 25 µg/mL. Note that 1X Diluted H9C2 and H4IIE cell extract samples were at 200 µg/mL The interpolated, dilution factor-corrected values are plotted in pg of IkappaB alpha per µg of total protein (mean +/- SD, n=2). As expected, treatment of NIH/3T3 cells with TNF-alpha reduces IkappaB alpha concentration.
To learn more about the advantages of recombinant antibodies see here.
Datasheets and documents
ab206308 has not yet been referenced specifically in any publications.