Apoptosis, or programmed cell death, is an essential process in the maintenance of tissue homeostasis and embryonic development. Its deregulation is implicated in several pathologies, such as neurodegenerative diseases and cancer. To advance research into these processes, it is crucial to have precise tools to detect and quantify the different stages of apoptosis. In this context, Immunostep offers a range of kits and tools specifically designed for the sensitive detection of apoptotic stages by flow cytometry.
Importance of apoptosis detection
Understanding the mechanisms of apoptosis is critical in areas such as cancer research, where resistance to cell death is a hallmark of tumor cells. Furthermore, in drug development, assessing the induction of apoptosis in treated cells is vital to determine the efficacy and safety of new compounds. Therefore, reliable methods to identify cells at different stages of apoptosis are indispensable for researchers and clinicians.
Immunostep Apoptosis Detection Kits
Immunostep has developed a series of kits that allow the accurate identification of apoptotic cells at different stages, from early to advanced stages. These kits are validated for use in flow cytometry with various cell types and drug treatments, offering fluorescent options adapted to multiple laser excitation sources.
Apoptotic membrane assays
A common method to detect apoptosis is staining with Annexin V, a protein that binds strongly to phosphatidylserine (PS). In healthy cells, PS remains on the inner side of the plasma membrane. During early apoptosis, PS is externalized, making it detectable by Annexin V conjugated with fluorochromes like FITC or PE. To differentiate apoptotic from necrotic cells, double staining is used. Dyes like propidium iodide (PI) or 7-ADD penetrate only membrane-compromised cells, indicating necrosis. This combination allows differentiation between viable, apoptotic and necrotic cells in a single sample.
Mitochondrial assays
Mitochondrial dysfunction is a prominent feature in the early stages of apoptosis. Immunostep’s MitoStep kits are designed to measure changes in mitochondrial membrane potential using the cationic dye DilC1(5). In cells with functional mitochondria, this dye accumulates, emitting intense fluorescence. During apoptosis, the loss of mitochondrial membrane potential results in a decrease in fluorescence, allowing the identification of cells in early stages of apoptosis. This method is especially useful for detecting early changes before alterations in the plasma membrane occur.
Research and Diagnostic Applications
Immunostep’s apoptosis detection kits are versatile tools applicable in multiple fields:
-Oncology research: evaluate the efficacy of anti-tumor therapies by quantifying apoptotic cells after specific treatments.
-Neurodegenerative diseases: study the mechanisms of cell death in pathologies such as Alzheimer’s or Parkinson’s, where apoptosis plays a crucial role.
-Drug development: to determine the cytotoxicity and mechanism of action of new pharmacological compounds, it is essential to evaluate their ability to induce apoptosis in target cells.
-Cell viability assessment: In cell and molecular biology studies, it is essential to distinguish between viable, apoptotic, and necrotic cells. This distinction allows for a concrete interpretation of the experimental results.
Advantages of Immunostep kits
By choosing Immunostep’s kits, researchers benefit from:
-High sensitivity: ability to detect subtle changes in the early stages of apoptosis.
-Versatility: Compatibility with a wide range of cell types and treatments,
-Ease of use: optimized protocols that facilitate laboratory implementation.
-Diverse fluorescent options: availability of different fluorescent conjugates adapted to the most common flow cytometry equipment.
Conclusion
In summary, Immunostep’s apoptosis detection kits provide reliable and accurate tools for the investigation of programmed cell death, facilitating significant advances in areas such as oncology, neurosciences and drug development. Their design and validation ensure consistent results, supporting the scientific community in understanding and manipulating apoptotic processes.