SuperSepTM Phos-tagTM (50μmol/L), 12.5%, 13well
- for Electrophoresis
- Manufacturer :
- FUJIFILM Wako Pure Chemical Corporation
- Storage Condition :
- Keep at 2-10 degrees C.
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- Structural Formula
- Label
- Packing
- SDS
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5EA
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In stock in Japan |
※Check availability in the US with the distributor.
Document
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. This is a precast gel added with Phos-tag™ Acrylamide in advance. It can be used immediately after opening the package. It contains zinc as a supplemental metal. It has excellent storage stability by its neutral gel buffer. Sharp bands can be obtained. Features:
Physical Properties:
Please note that prestain marker does not reflect the molecular weight, when a gel containing Phos-tag™ is use. It can only be used as reference of electrophoresis. Please use the result as an index of transcription efficiency at Western blotting. At Western blotting, Zn2+ must be sufficiently removed with transfer buffer containing 10 mM EDTA. Please wash out the gel for 10 minutes x 3 times. Subsequently, perform the same procedure for 10 minutes using transfer buffer without EDTA, then transfer the gel. Please contact us to obtain an adjuster for your tank. |
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Property
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.