This is a knockout-validated antibody summary, based on the publication "Priming Phosphorylation of TANK-Binding Kinase 1 by IκB Kinase β Is Essential in Toll-Like Receptor 3/4 Signaling", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).
Mouse monoclonal IgG2b κ
Company: Santa Cruz
Antibody: IKKα
Catalog number: sc-7606
Summary: Mouse monoclonal IgG2b κ against amino acids 1-745 representing full length human IKKα. Recommended for detection of IKKα of human origin by western blot, immunoprecipitation, immunofluorescence, immunohistochemistry (paraffin), flow cytometry and ELISA.
Western blot
WT, IKKα-KO, and IKKα/β-DKO 293T cells.
Tris-buffered saline with 0.1% Tween 20 (TBST) containing 5% skim milk for 30 min at room temperature.
Diluted in the blocking buffer overnight at 4°C.
Horseradish peroxidase (HRP)-conjugated secondary antibody in TBST for 1 h at room temperature.
LAS-4000 instrument (Fujifilm) by chemiluminescence using Chemi-lumi One Super (Nacalai Tesque).
Figure: WT 293T cells and IKKα, IKKβ, and IKKα/β KO 293T cells were transfected with expression constructs for HA-tagged TLR3 and Flag-tagged IRF-3 S386A. Twenty-four hours later, cells were stimulated with 50 μg/ml of poly(I·C) for 2 h. Cell lysates were subjected to immunoprecipitation with anti-HA, followed by immunoblotting. Please see Figure 4g in the article [1].
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