JAPAN USA CHINA


Price & Availability : Please Select your country

 

-SulfoBiotics- Protein Redox State Monitoring Kit

Item # Unit Size
SB11-10
5 samples
SB12-10
5 samples

For Research Use Only Products

  • Number of thiol group(s) can be monitored clearly in a simple procedure
  • Two different types of the product available depending on your experiment
(SB11)          (SB12)


Storage Condition : 0-5
°C
Shipping Condition : ambient temperature
Kit Contents :
There are 2 different products available. If you need to perform Western blotting, please choose "Protein Redox State Monitoring Kit Plus" (Code#: SB12).

[Code#: SB11] Protein Redox State Monitoring Kit
Protein-SHifter x 5
Reaction Buffer A x 1
Reaction Buffer B x 1

[Code#: SB12] Protein Redox State Monitoring Kit Plus
Protein-SHifter Plus x 5
Reaction Buffer A x 1
Reaction Buffer B x 1
Lysis Buffer x 1



Product Description:

Modification of free-thiol group(s) in a protein is one of the typical posttranslational modifications and such modification is controlled by in vivo redox responses. In recent studies, it has been investigated that the modified free-thiol group controls many of cellular responses, e.g. cell death, protein expression, transcription.

This kit is designed to monitor the number of thiol group in protein with very simple procedure, and the number of thiol group can be monitored by electrophoresis (Code#: SB11) and western blotting (Code#: SB12). 

                            Fig. 1 Redox States of Thiol group(s) in Protein




Principles:

Labeling Principle using Protein-SHifter (Code#: SB11)
Maleimido group of the Protein-SHifter binds to a free thiol group of protein to modify the molecular weight of protein.
This modification allows the number of free-thiol(s) in protein to be visualized in electrophoresis.



Labeling Principle using Protein-SHifter Plus (Code#: SB12)
Maleimido group of the Protein-SHifter Plus binds to a free thiol group of protein to modify the molecular weight of protein.
This modification allows the number of free-thiol(s) in protein to be visualized in western blotting.
Photocleavage procedure works on to achieve better membrane transfer efficiency in western blotting. 


Visualization of Thiol number(s)

Since one molecule of Protein-SHifter binds to one molecule of free-thiol in protein, the redox state of the protein can be monitored as clear band-shift according to the number of Protein-SHifter bound(=number of free-thiol).


Experimental Data:

Visualization of SH number in GAPDH using Protein-SHifter (Code#: SB11)

*GAPDH: Glyceraldehyde-3-phosphate dehydrogenase

Protein-SHifter labeled GAPDH was electrophoretically-fractionated




Visualization of SH number in GAPDH using Protein-SHifter Plus (Code#: SB12)

1. Control (non-treated) + Protein-SHifter Plus labeling 
2. Diamide oxidation + Protein-SHifter Plus labeling
3. H 2O2 oxidation + Protein-SHifter Plus labeling

Once treated with oxidants, HeLa cells were lysed. GAPDH in cell lysate were labeled with Protein-SHifter Plus. Then the labeled sample was applied for electrophoresis. After the western blotting, the number of thiol group in GAPDH were monitored as shown.