This is a knockout-validated antibody summary, based on the publications "Identification of TBK1 complexes required for the phosphorylation of IRF3 and the production of interferon β", as cited below [1], and "I-κB Kinase-ε Deficiency Attenuates the Development of Angiotensin II-Induced Myocardial Hypertrophy in Mice" for western blot knockout validation (figure 2a) [2]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. BMDM from WT, IKKε KO, OPTN (D477N) and OPTN (D/N) × IKKε KO mice were stimulated with 10 mg/ml poly(I:C) for the times indicated. Aliquots of the cell extracts were subjected to SDS–PAGE and immunoblotted with antibodies that recognize IKKε. Antibodies to GAPDH were used as a loading control. From [1].

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: IKKε

Catalog number: 3416

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues near the carboxyl terminus of mouse IKKε. Reacts with mouse and rat. Suitable for western blot, immunoprecipitation, immunofluorescence/immunocytochemistry and flow cytometry.

Western blot

Bone marrow-derived macrophages (BMDM) from WT and IKKε KO mice. Cells were rinsed in ice-cold PBS and extracted in ice-cold lysis buffer (50 mM Tris–HCl ( pH 7.5), 1 mM EGTA, 1 mM EDTA, 1% (v/v) Triton X-100, 1 mM sodium orthovanadate, 50 mM sodium fluoride, 5 mM sodium pyrophosphate, 0.27 M sucrose, 10 mM sodium 2-glycerophosphate, 1 mM phenylmethylsulfonyl fluoride, 1 mM benzamidine and 1 mM dithiothreitol).

ECL detection system (GE Healthcare).

TANK (Cat #2141), TRIF (Cat #4596) and TAK1 (TGFβ-activated kinase 1; Cat #4505) from Cell Signaling are also KO validated (Fig 2 and 5).