SuperSepTM Phos-tagTM (50μmol/L), 15%, 13well
- for Electrophoresis
- Manufacturer :
- FUJIFILM Wako Pure Chemical Corporation
- Storage Condition :
- Keep at 2-10 degrees C.
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5EA
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Features
- Ready-to-use, saves time
- Able to isolate phosphorylated proteins by the level of phosphorylation
- Good separation with sharp bands
Applications
Albumin Dephosphorylation
[Electrophoresis Buffer] Tris-Glycine SDS Buffer
[Electrophoresis Samples]
Lane1: Untreated Albumin
Lane2: Dephosphorylated Albumin
[Electrophoresis cConditions] 20 mA, 70 mins
[Staining] Quick CBB staining
[Destaining] Deionized wWater
Albumin (Cat. No. 010-17071) was dephosphorylated using alkaline phosphatase (NIPPON GENE CO., LTD., Cat. No. 319-02661). Band shift confirmed dephosphorylation of albumin.
Time-dependent Dephosphorylation of β-casein
[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.
Time-dependent Change in the level of phosphorylation
Procedure
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.
Results
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.
Electrophoresis of Control Protein
P10 %(left): SuperSep™Phos-tag™ (50 μmol/L), 10 %, 13Wells
P15 %(middle): SuperSep™Phos-tag™ (50 μmol/L), 15 %, 13Wells
A15 %(right): SuperSep™Ace, 15 %, 13Wells
Electrophoresis Conditions
30 mA/gel (constant), 60 mins
Sample
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.
Running Buffer
Staining
Quick CBB Plus (Cat. No. 174-00553)
Overview / Applications
| Outline | This product is for research use only. Do not administer it to human. Phos-tag™ is a pre-cast gel for electrophoresis made with polyacrylamide, and can be used to separate phosphorylated and non-phosphorylated proteins. The gel has a polyacrylamide concentration of 15%. |
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Manufacturer Information
Alias
For research use or further manufacturing use only. Not for use in diagnostic procedures.
Product content may differ from the actual image due to minor specification changes etc.
If the revision of product standards and packaging standards has been made, there is a case where the actual product specifications and images are different.