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Fluoro Thiol
Fluorescent Thiol Detection Kit - Detection of Thiols in tissue extracts /cell samples
 
Key Benefits:
  • Detection of reduced Thiols in cells or tissue extracts
  • Sensitive Fluorescent Assay
  • Detection of reduced Thiol levels in apoptosis, metabolism and oxidative stress
  • detection of reduced thiols in Bacterial, fungal, plant cells or other samples
  • Fluorescent 96 well Plate Reader Readout (excitation: 488nm and emission at 515-530 nm)
Introduction & Assay Principle
During the normal course of metabolism, oxygen is partly reduced as electrons leak out of the electron transport chain during respiration. These partially reduced oxygen species (ROS) can react with organic substances through non-catalytic means. Furthermore, ROS can be generated via endogenous enzyme systems like plasma NADPH oxidase, cytoplasmic xanthine oxidase and organelle sources e.g., cytochrome P-450. ROS have been implicated in regulating diverse cellular functions including proliferation, defense against pathogens, intra-cellular signaling, transcriptional activation and apoptosis. Elevation of ROS beyond the buffering capacity of the cell can lead to oxidative stress. Elevated ROS levels can lead to damage of DNA/RNA, proteins and lipids which may lead to apoptosis. Cells have developed several mechanisms to counter act elevated ROS levels such as a thiol reducing buffer composed of cellular thiol levels (glutathione and thioredoxion) for the maintenance of the reduction-oxidation (redox) state of the cell, and enzymes to remove ROS (catalase, superoxide dismutase and glutathione peroxidase) (1-2).
Reaction
Cell Technology’s Fluoro- Thiol kit detects Thiol levels in cells and tissue extracts. Hatsuo Maeda and co workers have developed  2,4-Dinitrobenzenesulfonyl fluorescein (dye) as a fluorescent specific probe for general thiol detection (3). The reaction scheme is outlined below.

  Thiol (reduced) + non-fluorescent Dye   ------->         fluorescent analog

  excitation: 488nm and emission at 515-530 nm

 

Kit contents (for 100 assays)
  1. Dye: 1 vial Dried  ..........................................Part# 4021 - Store at +2-8 C
  2. Lysis Buffer: 1 Bottle 25 mL..........................Part# 3053 - Store at +2-8 C
  3. GSH Standard: 3 vials dried..........................Part# 7015 -  Store at +2-8 C
The following kits are available:
Catalog 
 
Size
 
 Price (Eur)
FLTHIO100-2 100 Tests €395

Figure 1. Detection of glutathione (GSH) utilizing the Fluoro Thiol kit.

 

Figure 2.  Jurkat cells were incubated with 1 mM staurosporine (4) for 3 hours. After which thiol levels were quantified using Cell Technology’s Fluoro Thiol kit. The graph represents approximately 1 x 10 4 cells per reaction (n=3).