Thioredoxin Redox State Assay Kit f...

Thioredoxin Redox State Assay Kit for Serum

Product Information

Product Overview:

Thioredoxin is a small (12 kDa) redox active thiol protein that is present in prokaryotic and eukaryotic cells. Thioredoxin maintains a thiol redox pool of cells and is known to be oxidized in conditions of oxidative stress. Mammalian thioredoxin has 3 structural cysteines at positions 62, 69 and 72; and two cysteines at positions 32 and 35 that take part in redox reactions. Once oxidized this protein is re-reduced by thioredoxin reductase and utilizes NADPH as a reducing equivalent. Mammalian thioredoxin is oxidized in response to oxidative stress and can remain in various redox states. For example, a strong oxidant such as H2O2 can fully oxidize thioredoxin which can remain in various redox states as shown in Fig 2. The Thioredoxin Redox State Assay will determine the relative abundance of oxidized and reduced thioredoxin in cellular lysates, or tissue homogenates. The thiol group is derivatized with an Iodoacetic Acid (IAA) and the oxidized and reduced thioredoxin are separated based on their relative mobility in a native polyacrylamide gel. The oxidized and reduced bands are visualized by western analysis as demonstrated in Fig 1. The bands can be quantified by densitometric determination of relative intensity of oxidized and reduced bands. Fluorescent labeling can also be done for direct measurement of these bands using secondary antibody conjugated with appropriate fluorescent label.

Application:

Intracellular redox state is an important regulator of gene expression, signal transduction and antioxidant defense system. The redox state of thioredoxin is a key indicator of redox environment of the cell. In a reduced state thioredoxin can induce gene expression, activate transcription factors and can detoxify peroxides via peroxiredoxins. However, increased oxidative stress could oxidize thioredoxin resulting in loss of function that is dependent on reduced thioredoxin. Therefore, it is important to determine the relative levels of reduced and oxidized thioredoxin in cells or tissues in response to a stress. Thioredoxin redox state can also be used as a marker of oxidative stress in cancer research, cardiovascular research, neurodegenerative disease research, and in toxicological research. This assay along with our Thioredoxin Activity Assay Kit (Cat #TR-7011K) provides a complete understanding of the response of thioredoxin systems to a specific oxidant or other thioredoxin modulating stimuli.

Features:

1. Assay buffer
2. Iodoacetic Acid
3. G-25 Spin Columns
4. Thioredoxin Antibody sufficient for 10 mini gels.

Catalog#: RS-8011K2
LID#: 219A
Discontinued
Order by Phone or Fax: 1.800.316.3081
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Specifications

Required Equipment1. Electrophoresis apparatus
2. Electroblotting apparatus
3. Western blotting reagents
MSDS AvailabilityYes, upon request.
Unit Size20 REACTIONS
Country of OriginUnited States of America
Safety StatementsFor research use only. Not for use in diagnostic procedures.
Sample AvailabilityNo

Shipping & Handling

Estimated Ship Time: 3 Business Days

Shipped In: Dry Ice

Hazardous Material: No

Storage

Storage Temperature: -20C

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