SensoLyte® 520 Thiol Quantitation Kit – Ultra-Sensitive


Thiol compounds, such as glutathione (GSH), cysteine, and homocysteine, are a natural reservoir of the reductive capacity of a cell. They function as components of the intracellular and extracellular redox buffer and play important roles in a variety of biological processes, such as enzyme catalysis, redox-signaling protein folding, and free radical scavenging.1-3

AnaSpec is pleased to announce the release of the SensoLyte® 520 Thiol Quantitation Kit (patent pending), an ultra-sensitive, robust and easy-to-use fluorescent assay for measurement of total thiol level. This kit contains a novel fluorogenic reagent which upon reaction with thiol releases fluorescence. Fluorescence signal can be detected at the common green excitation/emission wavelengths of 490 nm/520 nm. The assay has a linear range of 0.04-160 mM thiol. Common contaminants, e.g. DMSO, and Triton X-100, are well tolerated in the assay. Additionally, autofluorescence interference from cellular components or from test compounds is minimal at the emission wavelength of 520 nm.


Figure 1. Detection of GSH with the SensoLyte® 520 Thiol Quantitation Kit.
Fluorescence signal was detected at Ex/Em=490/520 nm (FlexStation 384II).

AnaSpec also offers another fluorimetric kit, the SensoLyte® ABD-F Thiol Quantitation Assay Kit (Figure 2); as well as a colorimetric kit, the SensoLyte® Thiol Quantitation Assay Kit (Figure 3).

SensoLyte® 520 Thiol Quantitation Assay (Cat# 72138)

SensoLyte® ABD-F Thiol Quantitation Assay (Cat# 72137)

SensoLyte®
Thiol
Quantitation Assay (Cat# 72136)

Read-out

Fluorimetric

Fluorimetric

Colorimetric

Wavelengths

Ex/Em= 490/520 nm

Ex/Em= 389/513 nm

Abs=405 nm

Hydrolysis Product

A green fluorophore

ABD-F thiol adducts

2-Nitro-5-thiobenzoic acid (TNB)

Lowest Detection Limit

0.04 uM

0.25 uM

1.5 uM

Kit size

(96-well plate)

500 tests

500 tests

500 tests



Figure 2. GSH reference standard. Serial dilutions of GSH were mixed with the substrate solution from the SensoLyte® ABD-F Thiol Quantitation Assay Kit.Fluorescence signal was detected at Ex/Em=389nm/513nm (FlexStation 384II, Molecular Devices). The detection limit can reach as low as 0.25 mM of GSH.


Figure 3. GSH reference standard. Serial dilutions of GSH were mixed with thiol detection solution from the SensoLyte® Thiol Quantitation Assay Kit and absorbance was measured at 405 nm (Ultra Microplate Reader EL808, Bio-Tek Instruments, Inc). Detection limit can reach as low as 1.5
mM of GSH.

Related Products

AnaLyte™ OPA Protein Quantitation Kit *Fluorimetric*

References

1. Dickinson, DA. et al. Biochem. Pharmacol. 64, 1019 (2002).
2. Winterbourn, CC. et al. Free Radical Biol. Med. 45, 549 (2008).
3. Franco, R. et al. Arch. Physiol. Biochem. 113, 234 (2007).