Rapid Detection and Quantification Service of ER Stress Model
The actual endoplasmic reticulum stress is difficult to measure directly. In most cases, measuring ER stress requires the introduction of exogenous proteins or indirect detection of mRNA/protein levels. To be able to observe protein aggregation during ER stress, Creative Bioarray uses Thioflavin T (ThT) to detect and quantify endoplasmic reticulum stress in living cells. This method can provide our customers with direct quantitative measurement of ER stress in live cells in real-time data. This new tool will be useful for future studies on the role of protein misfolding in the development and/or progression of human diseases.
Technology Principle
Thioflavin T is a small molecule with fluorescent properties that have been shown to selectively bind protein aggregates, particularly β-folds. Upon binding to these structures, ThT exhibits a change in emission maximum (from 445 nm to 482 nm) and an increase in fluorescence intensity of several orders of magnitude. These properties make ThT a commonly used dye in vitro and in vivo for the detection, diagnosis, and analysis of amyloidogenic fibrils and their associated pathologies in brain tissue and pancreatic islets. Enhanced ThT fluorescence correlates directly with established indicators of activation of the unfolded protein response.
Rapid Detection of Endoplasmic Reticulum Stress in Cells
- Cell types for which services are available include but are not limited to
Mouse embryonic fibroblasts (MEF), human hepatocellular carcinoma (HepG2) cells, human aortic endothelial cells (HAEC)
- Fluorescence detection and quantification
In addition to cells suspected of being in a state of oxidative stress in the endoplasmic reticulum, customers will be asked to provide cells in a non-stressed state as a control. We will first perform a pre-experiment to determine whether the cells to be tested are ER-stressed in response to ThT fluorescence and determine the concentration of ThT required to provide a significant and detectable signal.
We will perform appropriate cultures of the cells to be tested provided by the customer. Images of live cells will be captured and fluorescence quantified after adding the appropriate concentration of ThT to the culture.
Rapid Detection of Endoplasmic Reticulum Stress in Tissues
Freshly frozen tissue provided by the client is collected on coverslips and stored at -20°C. Sections are allowed to thaw prior to ThT staining and then fixed in 37% formaldehyde at room temperature. Sections were then washed with water and incubated with freshly filtered ThT in PBS at room temperature.
Service Features
- Enhanced ThT-fluorescence can be detected in living cells within 20 min of application of endoplasmic reticulum stress inducers.
- Thioflavin T-fluorescence is a direct measure of endoplasmic reticulum stress.
- This method can be used to continuously monitor/quantify ER stress in living cells.
- The method can be applied to a wide range of cell types and ER stress inducers.
- The method can be used to detect ER stress in tissue samples.
Creative Bioarray is dedicated to providing high-quality products, comprehensive services, and tailored solutions to support and facilitate life sciences and pharmaceutical research and development. If you have any questions or needs, please contact us, and our customer service staff will help you the first time.
All services and products are for scientific use only, not for medical use！
