Albumin (Cat. No. 010-17071) was dephosphorylated using alkaline phosphatase (NIPPON GENE CO., LTD., Cat. No. 319-02661). Band shift confirmed dephosphorylation of albumin.

Electrophoresis Buffer: Tris-Glycine SDS buffer
Electrophoresis Samples:
　Lane1: β-casein (AP-treatment: 0 min)
　Lane2: β-casein (AP-treatment: 15 min)
　Lane3: β-casein (AP-treatment: 30 min)
　Lane4: β-casein (AP-treatment: 45 min)
　Lane5: β-casein (AP-treatment: 60 min)
Electrophoresis Conditions: 35mA, 60min
Staining: Quick CBB staining
Destaining: Deionized Water

Dephosphorylation of β-casein was performed over different time periods. The results confirmed separation of phosphorylated and dephosphorylated β-casein.

In addition, the results demonstrated the level of dephosphorylation across different time points.

Data was provided by courtesy of Dr. Atsushi Enomoto, Molecular Radiology / Sec. of Radiation Biology, Center for Disease Biology and Integrative Medicine, Faculty of Medicine, University of Tokyo

X-ray irradiation (5 Gy) was performed for human lung cancer cell line Lu99, and cells were collected across different time points. Cell extracts were prepared, and SDS-PAGE was performed in 13 wells using 50 μmol/L of 10% SuperSep Phos-tag.

Gentle agitation of the gel was performed in a transfer buffer containing 10 mmol/L EDTA, and proteins were transferred to PVDF membrane. The membrane was blocked with 2% Milk/TBS-T, and was reacted with primary antibodies (upper image: p53, lower image: cell-cycle proteins). Detection was performed by chemoluminecence.

Protein accumulation for p53 was at the highest level 4 hours after X-ray irradiation. The level of protein X phosphorylation changed over time after X-ray irradiation.

P10 % (left): SuperSepPhos-tag (50 μmol/L), 10%, 13Wells
P15 % (middle): SuperSepPhos-tag (50 μmol/L), 15%, 13Wells
A15 % (right): SuperSepAce, 15%, 13Wells

30 mA/gel (constant), 60 mins

5 μg/lane of α-casein from bovine milk, dephosphorylated (Cat. No. 038-23221)

The product contains α-casein that hasve not been dephosphorylated. While conventional SDS-PAGE will only show a single main band, Phos-tag SDS-PAGE will show two main bands for α-casein and dephosphorylated α-casein.

Quick CBB Plus (Cat. No. 174-00553)