Anti-AKT1 (phospho-S246) Antibody

Affinity-purified by Antibodies.com, this rabbit polyclonal antibody specifically recognizes AKT1 only when serine 246 in the linker region is phosphorylated, allowing researchers to monitor PI3K/AKT pathway activation rather than total AKT1 levels. The unconjugated reagent is supplied at 1 mg/ml in a 100 µl vial containing phosphate buffered saline with 0.05% sodium azide at approximately pH 7.2, and is compatible with standard HRP- or fluorophore-conjugated secondary antibodies.

Validated applications include Western blot and immunohistochemistry on human, mouse, and rat samples, covering the species most frequently used in cancer and metabolism research. S246 phosphorylation reflects AKT1 autophosphorylation after membrane recruitment, so lysates from insulin- or EGF-stimulated cells provide a useful positive control, while λ-phosphatase treatment of matched samples confirms phospho-specificity.

For Western blot, the antibody is generally used at 0.5–2 µg/ml (roughly 1:500 to 1:2000 from the 1 mg/ml stock) and produces a band near 56–60 kDa corresponding to endogenous AKT1. For IHC on formalin-fixed paraffin-embedded tissue, citrate buffer heat-induced epitope retrieval at pH 6 is recommended. The 100 µl volume supports several full Western blot experiments or dozens of stained slides, depending on dilution. Store refrigerated at 4°C; for extended storage, divide into smaller aliquots and keep frozen at -20°C, avoiding repeated freeze-thaw cycles that erode phospho-epitope recognition. The 0.05% sodium azide acts as a preservative in working solutions.