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Applications Table
| Western Blot | Chromatin IP | Immunoprecipitation |
|---|---|---|
| 1 - 2 µg/ml | — | 1 - 2 µg/ml |
for IKKa antibody (mAb)Background
IKKα (IκB Kinase alpha subunit) – The IκB Kinase (IKK) complex is the key enzyme involved in the activation and translocation of NFκB (NFκB p50 and NFκB p65). The IKK complex is composed of two catalytic subunits (IKKα and IKKβ) and a regulatory subunit (IKKγ).
NFκB signaling is negatively regulated by the sequestration of the NFκB complex in the cytoplasm by its association with the IκB family of inhibitory proteins. Upon its activation, the IKK complex phosphorylates IκBα at Ser32 and Ser36. This results ultimately in the degradation of IκB and the activation and nuclear translocation of NFκB. While both IKK-alpha and IKK-beta subunits are Ser/Thr kinases, IKK-beta appears to be the principal kinase, as IKKα is not required for activation of the IKK complex and the subsequent degradation of IκBα.
for IKKa antibody (mAb)Immunogen
This antibody was raised against a full-length protein corresponding to human IKKα.
for IKKa antibody (mAb)Application Notes
This IKKα antibody has been validated for use in Western blotting (1 - 2 µg/ml dilution) and immunoprecipitation (1 - 2 µg/ml dilution). For optimal results, primary antibody incubations should be performed at room temperature. The addition of 0.1% Tween 20 to all Blotto solutions may also reduce background. Individual optimization may be required.
for IKKa antibody (mAb)Buffer
PBS containing 0.02% sodium azide. Sodium azide is highly toxic.
for IKKa antibody (mAb)Positive Control
Active Motif's Daudi nuclear extract (Catalog No. 36091) can be used as a positive control for IKKα antibody.
Bridging Antibody to Improve ChIP, MeDIP and IP
Some isotypes of mouse IgG antibodies do not bind well to protein G-conjugated agarose/magnetic beads, which are commonly used in ChIP, MeDIP and IP experiments. Capture efficiency can be improved by including an anti-mouse IgG bridging antibody because it binds with a strong affinity to both the protein G beads and the mouse IgG antibody. By increasing the affinity of the mouse primary antibody for the protein G bead, the Bridging Antibody for Mouse IgG can help improve the results of all ChIP, MeDIP and IP experiments that use IKKα antibody (mAb).
IKKα mAb tested by Western blot.
IKKα detection by Western blot analysis. The detection of IKKα was performed using 30 µg of whole-cell extract from Daudi cells with the IKKα mAb at a 1 µg/ml dilution.
for IKKa antibody (mAb)Storage
Antibodies in solution can be stored at -80°C for 2 years. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.
for IKKa antibody (mAb)Guarantee
This product is guaranteed for 6 months from date of receipt.
This product is for research use only and is not for use in diagnostic procedures.
Application Key
- ChIP = Chromatin Immunoprecipitation;
- DB = Dot Blot;
- ELISA = Enzyme-linked Immunosorbent Assay;
- EMSA = Electrophoretic Mobility Shift Assay
- FACS = Flow Cytometry;
- ICC = Immunocytochemistry;
- IF = Immunofluorescence;
- IHC = Immunohistochemistry;
- IP = Immunoprecipitation;
- MeDIP = Methyl-DNA Immunoprecipitation;
- WB = Western Blotting


