Detection of Human Phospho-p27/Kip1 (T198) by Western Blot.
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.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
The cyclin-dependent kinase (cdk) inhibitor p27/Kip1 is a tumor suppressor protein that acts in the nucleus to enforce cell cycle checkpoints. p27/Kip1 binds to and inhibits many cyclin/cdk complexes, including cyclin D/cdk4, cyclin E/cdk2, and cyclin A/cdk2. This inhibition can subsequently block progression through different phases of the cell cycle. p27/Kip1 is phosphorylated at T198 in cells treated with the AMP-dependent protein kinase (AMPK) activator AICAR (5-amino-4‑imidazolecarboxamide riboside) or glucose withdrawal.
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