Cell Apoptosis Assessment Services
Introduction to Cell Apoptosis and Associated Assays
Apoptosis, commonly known as programmed cell death, is a vital process in the maintenance of cellular homeostasis, development, and elimination of damaged or diseased cells. The intricate cascade of cellular and molecular events during apoptosis ensures that cells die in a controlled manner, without eliciting an inflammatory response. Dysregulation of apoptosis is associated with various diseases, including cancer, neurodegenerative disorders, and autoimmune diseases. Therefore, studying apoptosis is crucial in biomedical research and therapeutic development.
Fig.1 Stages of apoptosis.1
To study apoptosis, a range of assays have been developed, each targeting specific features of the apoptotic pathway. These include TUNEL assays for detecting DNA fragmentation, Annexin V staining for identifying phosphatidylserine exposure on the cell membrane, caspase activity assays that measure the activation of proteases central to the execution of apoptosis, etc.
Our Apoptosis Assay Services
At Creative Biolabs, we provide an extensive range of state-of-the-art flow cytometry (FC)-based apoptosis assay services tailored to various aspects of apoptosis evaluation. Our services include:
Annexin V Apoptosis Assay
The Annexin V Apoptosis Assay is a gold standard for detecting early apoptotic cells. Using a fluorophore-conjugated Annexin V, we quantitatively measure the percentage of apoptotic cells. Coupled with Propidium Iodide (PI) staining, this assay differentiates early apoptotic cells (Annexin V+/PI-) from late apoptotic/necrotic cells (Annexin V+/PI+).
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ATM and H2A.X Assay
Ataxia telangiectasia mutated (ATM) and phosphorylated H2A.X (γH2A.X or activated H2A.X) are key markers of DNA damage response. Our FC-based ATM and γH2A.X assay is used to assess activations of ATM and H2A.X during apoptosis. By combining flow cytometry with specific antibodies against these markers, we efficiently measure and analyze DNA damage responses at a single-cell level.
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Caspase Activation Assay
Caspase activation is a hallmark of apoptosis induction. Our FC-based caspase activation assay enables the accurate detection and quantification of active caspase enzymes, providing a detailed and quantifiable assessment of apoptotic events. Our assay leverages specific fluorescent substrates that, when cleaved by active caspases, emit a measurable signal.
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DNA Fragmentation Assessment
Flow cytometry is a powerful tool used to assess DNA fragmentation, which is a hallmark of apoptosis. Several methods are employed in FC-based DNA fragmentation assessment to detect and quantify apoptotic cells, including the TUNEL assay, Hoechst staining assay, and DAPI staining assay.
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H2A.X Assay
The phosphorylated form of H2A.X appears at sites of DNA double-strand breaks, an event linked to apoptotic processes. By using FC, we effectively assess the total H2A.X and γ-H2A. H2A.X assay allows for the sensitive and rapid detection of DNA damage and repair processes. This assay is particularly useful in studies of DNA damage and repair, apoptosis, and cellular responses to genotoxic stress.
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Mitochondrial Depolarization Assessment
Mitochondrial depolarization typically occurs during the early stages of apoptosis. Specific mitochondrial dyes, such as JC-1 or TMRM, accumulate within the mitochondria under normal, polarized conditions. When mitochondrial membrane integrity is compromised—a hallmark of apoptotic progression—these dyes leak out, resulting in quantifiable changes in fluorescence that can be detected and measured by FC.
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Reference
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Chmurska, Anna, Karolina Matczak, and Agnieszka Marczak. "Two faces of autophagy in the struggle against cancer." International Journal of Molecular Sciences 22.6 (2021): 2981. Distributed under Open Access license CC BY 4.0, without modification.
For Research Use Only | Not For Clinical Use
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