It has been added to the cart.

It has been added to the comparison table.

SuperSepTM Phos-tagTM (50μmol/L), 10%, 13well

for Electrophoresis
Manufacturer :
FUJIFILM Wako Pure Chemical Corporation
Storage Condition :
Keep at 2-10 degrees C.
  • Structural Formula
  • Label
  • Packing
SDS
Comparison
Product Number
Package Size
Price
Inventory
Distributor
193-16711
Barcode No
4548995006288
5EA
List Price
330.00 USD

In stock

Document

SDS
Product Specification Sheet
Spectral Data
Certificate of Analysis
Calibration Certificate

Features

00908_img02.gif
  • Ready-to-use, saves time
  • Able to isolate phosphorylated proteins by the level of phosphorylation
  • Good separation with sharp bands

Applications

Albumin Dephosphorylation

00908_img03.jpg

[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

00908_img04.jpg

[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

00908_img05.png
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

00908_img06.jpg

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 product is a pre-cast gel for electrophoresis made with polyacrylamide.
The Phos-tag contained in a gel during SDS-PAGE supplements the phosphoproteins, retarding the electrophoretic velocity of the phosphoproteins. In this way, the phosphoproteins and unphosphorylated proteins can be detected as separate bands.

Property

Manufacturer Information

Alias

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.

877-714-1920

Hours of Operation: 8:00 - 17:00 (EST)For other hours than the above, please contact us via the inquiry form.