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SuperSepTM Phos-tagTM (50μmol/L), 12.5%, 13well

for Electrophoresis
Manufacturer :
FUJIFILM Wako Pure Chemical Corporation
Storage Condition :
Keep at 2-10 degrees C.
  • Label
  • Packing
SDS
Comparison
Product Number
Package Size
Price
Availability
Certificate of Analysis
Purchase
Distributor
195-16391
Barcode No
4987481627536
5EA
List Price
330.00 USD

In stock in Japan

Document

SDS
Product Specification Sheet
Certificate of Analysis

Features

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  • Ready-to-use, saves time
  • Able to isolate phosphorylated proteins by the level of phosphorylation
  • Good separation with sharp bands

Applications

Albumin Dephosphorylation

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[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

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[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

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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
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

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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:

  1. Phosphorylated- and non-phonphorylated proteins are efficiently separable with sharp bands.

  2. Long term stability (stable for 6 months)

Physical Properties:
  • Plate size: 100 x 100 x 3 (mm)

  • Gel size: 90 x 85 x 1 (mm)

  • Well number: 13

  • Well volume: 30 μL

  • Note:
    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.

    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.