Western blot shows lysates of MCF‑7 human breast cancer cell line, non-transfected, transfected with plasmid expressing YFP-p27/Kip1 wild type (WT), or transfected with YFP-p27/Kip1 T198A mutant (T198A). PVDF membrane was probed with 0.5 µg/mL Rabbit Anti-Human Phospho-p27/Kip1 (T198) Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3994) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog # HAF008). A specific band for p27/Kip1 was detected at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.