Quantification of Free Sulfhydryl Groups in Rat Cells

Free sulfhydryl groups, mainly refer to biomolecules containing sulfhydryl groups (-SH), exist in cells in the form of glutathione (GSH). GSH is one of the most abundant antioxidants in cells, it can remove harmful oxidants and maintain the redox balance in cells. Abnormalities in free sulfhydryl groups, such as their decreased concentrations, are often associated with the development of various diseases, including cancer, neurodegenerative diseases, and cardiovascular diseases. In experimental animal models, especially in rat models, the quantitative analysis of free sulfhydryl groups can be used to evaluate the toxic effects of drugs, monitor the progression of diseases, and study the effects of antioxidant treatments. For example, in the study of a new anticancer drug, scientists can evaluate the oxidative stress response of the drug by measuring the GSH level in rat tumor cells.

 

Methods for Determining Free Sulfhydryl Groups

In rat cells, free sulfhydryl groups are usually measured by the following techniques:

 

1. Chromatography

High-performance liquid chromatography (HPLC) combined with fluorescent probes can accurately measure the concentration of GSH in cells.

 

2. Electrochemistry

The sulfhydryl content in cell samples can be directly measured by an electrochemical sensor, which responds quickly and has high sensitivity.

 

3. Ellman's Method

Using specific reaction reagents (such as 5,5'-dithiobis-2-nitrobenzoic acid, DTNB), they can react with sulfhydryl groups to form a yellow compound, the absorbance spectrum of which can be measured to estimate the concentration of sulfhydryl groups.

 

Ellman's method is a routine method for detecting free sulfhydryl groups concentrations. The principle is that the DTNB reagent reacts with free sulfhydryl groups to form 2-nitro-5-thiobenzoic acid (TNB-), which can ionize in water under neutral or alkaline pH conditions to form TNB2- dianions. This TNB2- ion appears yellow, and the absorbance of visible light at 412nm can be measured to quantify TNB2-, then the free sulfhydryl groups can be quantified by the method of quantifying sulfhydryl groups with cysteine standards or based on molar absorbance.

 

MtoZ Biolabs has established seven detection platforms. Among them, the free sulfhydryl group content detection platform based on Ellman's method has complete method development and transfer capabilities.

 

Service Advantages

1. High Sensitivity

It can detect extremely low concentrations of free sulfhydryl groups, suitable for research on the minute changes of free sulfhydryl groups in cells.

 

2. High Selectivity

By selecting specific probes and separation conditions, specific types of free sulfhydryl groups can be quantitatively determined, avoiding the interference of other substances.

 

3. Real-Time Monitoring

Especially with fluorescent probe technology, the dynamic changes of free sulfhydryl groups in cells can be monitored in real time without destroying the cells.

 

4. Wide Range of Applications

Suitable for various biological samples, including cell culture, tissue sections, and animal models, etc.

 

For more service details, please feel free to consult!