Anti-IκBα Antibody

IKK beta (IκB Kinase, IKKβ, I-Kappa-B kinase-beta) is a member of the IKK complex which is composed of IKK alpha, IKK beta, IKK gamma and IKAP. IκB Kinase (IKK) complex is a critical component in NFκB regulation. In a majority of unstimulated cells, the NFκB transcription factors exist in their inactive form and are retained in the cytoplasm by the bound inhibitory IκB proteins. For NFκB to become activated, it must first disassociate from the inhibitor IκB. This occurs via the serine phosphorylation of IκB on Ser32 and Ser36. Post receptor stimulation, most notably by TNFα, NIK, a serine/threonine kinase, activates and phosphorylates IKKα and IKKβ. Phosphorylation of 2 sites at the activation loop of IKK beta is essential for activation of IKK by TNF and IL1. Once activated, IKK beta autophosphorylates, which in turn decreases IKK activity and prevents prolonged activation of the inflammatory response. Additionally, IKK beta activity can also be regulated by MEKK-1. IKK then binds to and phosphorylates IκBα on the two serine residues. This phosphorylation marks IκB for ubiquination and destruction by the proteosome. This frees the NFκB complex and allows it to translocate to the nucleus.

Recognizes IκBα, C-terminus. May react non-specifically with other proteins. Control peptide provided will compete only with the specific reaction of antiserum with IκBα.

Epitope: C-terminus

IκBα peptide corresponding to a region near the C-terminus of the human protein, conjugated to KLH.

Detect IκBα using this Anti-IκBα Antibody validated for use in WB, IH(P), ELISA & IP.

Immunohistochemisty (paraffin): Representative testing from a previous lot.
Anti-IкBα staining on invasive ductal carcinoma tissue (Breast Cancer) was pretreated with citrate buffer, pH 6.0. A 1:1,000 diluted was used using IHC-Select detection with HRP-DAB.


ELISA: Recommended

Immunoprecipitation: Recommended

Optimal dilutions must be determined by the end user.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Transcription Factors

Routinely evaluated by Western Blot on HEK293 lysates.

Western Blot Analysis:
1:500 to 1:2,000 dilution of this lot detected IkBalpha on 10 μg of HEK293 lysates.

~36 kDa

Replaces: AB3016

Rabbit polyclonal IgG serum in buffer containing 0.02 M Potassium phosphate with 0.15 M NaCl, pH 7.2 and 0.1% sodium azide.

Both antibody and control are stable for 6 months at -20ºC in undiluted aliquots from date of receipt.
Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.

Control
HEK293 cell lysate.
Control Peptide: Included with the antibody is 50 μg (1 mg/mL) of IκBα control peptide. The peptide will block the specific interaction of AB3016 with the IκBα subunit. Control peptide should be used at 1.0 μg per 1.0 μL of antiserum used in assay. Optimal concentrations must be determined by the end user.

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.