Anti-CHUK (phospho T23) Antibody

Raised in rabbit against a synthetic phosphopeptide corresponding to residues flanking threonine 23 of human IKK-alpha (CHUK), this unconjugated polyclonal IgG reagent from Antibodies.com selectively recognizes the kinase only when phosphorylated at this regulatory site. Phosphorylation of CHUK at Thr23 is mediated by AKT downstream of TNF receptor signaling, as first demonstrated by Ozes and colleagues, marking engagement of the canonical NF-kappa-B activation cascade through I-kappa-B-alpha degradation.

The affinity-purified antibody is supplied as 200 µl of liquid in PBS (pH 7.4) containing 150 mM NaCl, 50% glycerol, and 0.02% sodium azide, and should be stored at -20 °C with aliquoting recommended to limit freeze-thaw cycles. It reacts with human, mouse, and rat samples and is validated for Western blot at 1:500 to 1:1000 and immunohistochemistry at 1:50 to 1:100 on paraffin-embedded sections.

Working applications include tracking TNF-, IL-1-, or EGF-driven pathway activation in cell lysates (for example, MDA-MB-435 cells treated with 200 ng/ml EGF for 5 minutes serve as a positive Western blot control), localizing active kinase within tumor or inflamed tissue by IHC (human colon carcinoma is a suggested reference tissue), and parallel blocking experiments using the non-phosphorylated peptide to confirm phospho-specificity of the observed band or staining pattern.

Predicted band size is approximately 84 kDa, matching full-length CHUK. The polyclonal format delivers multi-epitope coverage around the T23 phosphosite, supporting robust detection across varied lysate preparations and across both formalin-fixed paraffin-embedded and frozen tissue sections used in standard signaling studies.