Maximize your apoptosis analysis
Apoptosis is one of the major forms of cell death and is associated with numerous diseases and biological processes; over >30,000 scientific publications a year mention apoptosis. As such, ensuring accurate measurement of apoptosis markers is essential for understanding disease mechanisms and the actions of drugs. At Abcam, we offer a comprehensive range of tools to measure apoptotic pathways effectively. From TUNEL assays to caspase activity detection, our products are designed to provide you with precise, reliable results every time. Whether you're focused on Cleaved Caspase 3, Cytochrome C, or Caspase 7, 8, and 9, we have the solutions for your apoptosis experiments.
Our apoptosis assay kits and reagents cover all the major stages of apoptosis, from loss of membrane asymmetry to DNA fragmentation. They employ simple methods that are widely accepted and have been used in thousands of publications. The readout of the assays can be performed using the most commonly available types of flow cytometers, plate readers, fluorescence microscopes, and other common instrumentation. Our apoptosis detection products feature TUNEL staining kits, Annexin V staining solutions, and a variety of Caspase activity assays, antibodies, recombinant proteins, and more. Each tool in our range is engineered to support your research in monitoring apoptosis pathways with unmatched accuracy. Whether your focus is on early-stage detection using Annexin V staining or later-stage apoptosis markers such as cleaved caspase 3, our products enhance your ability to track the progression and mechanism of apoptosis.
Discover the difference that our apoptosis assay products can make in your research. Browse our comprehensive selection today and find the perfect solution to measure key apoptotic markers and pathways. With a wide range of detection kits, including TUNEL assays, caspase assays, and Cytochrome C measurement tools, we’re here to support your research at every step.
For a deeper dive into cell health, including the background on methods to assess cell viability, cytotoxicity, mitochondrial function, the roles of the various caspases, and more, download our Cell Health Guide.
Annexin V assays
One of the earliest stages of apoptosis is loss of membrane asymmetry, where phosphatidylserine, which is normally restricted to the intracellular side of the plasma membrane, migrates to the outer plasma membrane.
Annexin V is a mammalian protein that binds to phosphatidylserine. Fluorescent dye conjugated Annexin V is commonly used to detect phosphatidylserine, using flow cytometry and fluorescence microscopy. The dye-conjugated Annexin V only binds to cells in which phosphatidylserine has migrated to the outer plasma membrane.
Our Annexin V FITC apoptosis staining kit (ab14085) is our most popular Annexin V assay. It includes an optimized binding buffer, propidium iodide to detect dead cells that have lost their membrane integrity, and a comprehensive protocol. Cells are resuspended in binding buffer, with a single staining step of 5 min, followed by detection using a flow cytometer or fluorescence microscope.
Product highlights
Annexin V-FITC Apoptosis Staining / Detection Kit
- Used in >500 scientific publications since 2006
- Convenient and complete apoptosis assay kit
10X Annexin V/ANXA5 Binding Buffer
- Optimized Annexin V binding buffer used in ab14085
- Suitable for fluorescent dye, fluorescent protein, and biotin Annexin V conjugates
Annexin V/ANXA5-FITC Apoptosis Detection Reagent ()
- Annexin V FITC conjugate used in ab14085
TUNEL assays and DNA fragmentation
DNA fragmentation is a late-stage apoptosis marker and is commonly detected using either by isolation of DNA and observation of fragments using an agarose gel or a TUNEL assay. TUNEL assays are a widely used method for detecting DNA fragmentation, a characteristic indicator of apoptosis. In this technique, a terminal deoxynucleotidyl transferase enzyme incorporate labeled deoxynucleotides at the ends of fragmented DNA strands. Alternatively, apoptotic cells can be identified by measuring cells with reduced DNA content using flow cytometry, employing a fluorescent dye such as propidium iodide. This approach enables the detection of cells that have lost DNA fragments, thus allowing for the identification of apoptotic cells with high specificity. DNA fragmentation occurs as a result of the activation of nucleases during apoptosis, leading to the formation of double-strand breaks. By detecting these breaks, TUNEL assays allow researchers to track the extent and progression of apoptosis in various cell populations.
TUNEL assays are especially valuable for monitoring the late stages of apoptosis, where DNA fragmentation becomes more prominent. It can be used on a range of samples, including tissues and cell cultures, making it a versatile means of detecting late-stage apoptosis. When paired with other apoptosis markers, such as caspase activity or membrane alterations, TUNEL staining provides a comprehensive picture of cell death and its underlying mechanisms.
Our TUNEL assay kits are suitable for use with paraffin-embedded tissue sections, frozen tissue sections, adherent and suspension cell cultures, and with preparations of single cell suspensions fixed on slides. Detection is performed by conventional light microscopy, fluorescence microscopy, or flow cytometry.
Product Highlights
Apoptosis DNA Ladder Assay Kit
- Kit for purification of fragmented DNA from cultured cells
- Simple analysis - run purified DNA fragments on am agarose gel
TUNEL Assay Kit – FITC
- Suitable for suspended and adherent cell cultures
- Analysis by flow cytometer or fluorescence microscope
- Supplied with TUNEL-positive and TUNEL-negative control cells
TUNEL Assay Kit – HRP-DAB
- Suitable for frozen and paraffin-embedded tissue sections, and cells coated on slides
- Analysis by conventional light microscopy
- Used in >200 scientific publications
Caspase assays
Caspases are a family of proteases central to the execution of apoptosis. These enzymes include Caspase 3, 7, 8, and 9, which play key roles in both the intrinsic and extrinsic apoptotic pathways. Caspase assays typically involve using substrates that become fluorescent or colorimetric upon cleavage by active caspases, allowing researchers to quantify apoptosis in real-time. By detecting the activity of specific caspases, these assays can help clarify which pathways are involved in apoptosis within a given cell population.
Cleaved caspase 3, in particular, is often used as a marker of apoptosis, as it signifies the irreversible commitment of a cell to die. Caspase-3 is activated through its cleavage by Capase 8, Caspase 9, or granzyme B. Cleaved caspase-3 can then be detected using assays based on:
- antibodies specific for cleaved caspase-3
- caspase enzyme substrates
- analysis of proteins cleaved by active caspase-3, namely PARP-1.
Other caspases, such as caspase 8 and caspase 9 can help determine whether apoptosis was initiated through external signals (extrinsic apoptosis) or mitochondrial pathways (intrinsic apoptosis). Caspase assays are essential for dissecting these pathways and gaining a better understanding of the molecular events leading to programmed cell death.
Our range of Caspase assays and products include ELISAs, antibodies for western blotting and imaging, recombinant proteins, and substrate kits suitable for measurement using plate readers and flow cytometers. Our tools of tools allow researchers to obtain a comprehensive view of Caspase activation in apoptosis.
Product highlights
Anti-Cleaved Caspase-3 antibody [E83-77]
- Suitable for Western blotting and fluorescence microscopy using human samples
- Used in >600 scientific publications
Caspase-3 Assay Kit (Colorimetric)
- Cleavage of DEVD-pNA substrate releases p-NA, with readout on colorimetric plate reader
- DEVD substrate also detects Caspase-7 activity
- Used in >300 scientific publications
Cleaved Caspase-3 Staining Kit (FITC)
- FITC-DEVD-FMK Caspase inhibitor binds to Caspases-3 and -7
- Detection by fluorescence microscopy, flow cytometry, or fluorometric plate reader
Mitochondrial membrane potential-dependent dyes
Mitochondrial membrane potential (Δψm) plays a crucial role in apoptosis, and measuring changes in this potential provides insight into mitochondrial dysfunction, a key event in the intrinsic apoptotic pathway. Mitochondrial membrane potential-dependent dyes are widely used to monitor this change. These dyes accumulate in mitochondria based on the membrane potential, and when the membrane potential collapses, indicating the initiation of apoptosis, the dye either fails to accumulate or diffuses out, and the fluorescence of the dye can then be measured. By detecting this loss of mitochondrial potential, researchers can observe early apoptotic events before other markers, such as DNA fragmentation or caspase activation, become evident.
Using these dyes helps identify cells undergoing apoptosis in the early stages, providing a deeper understanding of the mitochondrial involvement in programmed cell death. This is particularly useful for studies focusing on mitochondrial health, drug toxicity, and neurodegenerative diseases, where apoptosis plays a significant role. However, loss of mitochondrial membrane potential is not solely a marker of apoptosis; it is also associated with other types of cell death.
Abcam offers a comprehensive range of assays, tools, and reagents for the study of mitochondrial health and function.
Product highlights
TMRE-Mitochondrial Membrane Potential Assay Kit
- Suitable for fluorometric microplate readers, fluorescence microscope, and flow cytometry
- TMRE dye is sequestered in mitochondria due to its relative negative charge, depolarized mitochondria do not sequester the dye
JC-10 Mitochondrial Membrane Potential Assay Kit (Microplate)
- JC-10 forms orange aggregates as the mitochondrial membrane potential increases.
- Signal shifts from green (unaggregated) to orange (aggregated)
JC-10 Mitochondrial Membrane Potential Assay Kit (Flow Cytometry)
- Same JC-10 dye, with buffers formulated for flow cytometry
Cytochrome C release assays
Cytochrome C release from the mitochondria is a crucial event in the intrinsic apoptotic pathway. Once released into the cytosol, Cytochrome C interacts with apoptotic protease activating factor-1 (Apaf-1) and initiates the activation of caspases, particularly caspase 9. Cytochrome C release assays are used to measure this key step, providing insights into mitochondrial integrity and the onset of apoptosis. These assays typically involve isolating cytosolic and mitochondrial fractions from cells and detecting the presence of Cytochrome c through methods like western blotting or ELISA.
By tracking the release of Cytochrome C, researchers can pinpoint when and how mitochondrial damage occurs, offering a detailed look at the initiation of the apoptotic process. These assays are essential for studying the intrinsic apoptotic pathway, particularly in cancer research, neurodegenerative diseases, and toxicology analysis, where mitochondrial dysfunction often plays a central role.
We provide a full set of cytochrome antibodies suitable for western blotting and fluorescence microscopy, kits containing antibodies and the other reagents needed to successfully stain for Cytochrome C, and a Cytochrome C recombinant protein.
Product highlights
Cytochrome c Apoptosis ICC Antibody Kit
- Convenient set of antibodies and buffers to stain Cytochrome c in fluorescence cell imaging
- Comes with an antibody against Complex Va to stain the mitochondria
Cytochrome c Release Assay Kit
- Complete kit for purification of the cytosolic and mitochondrial fractions
- Supplied with a primary antibody to measure Cytochrome C in Western blotting
Anti-Cytochrome C antibody [EPR1327]
- Recombinant anti-Cytochrome C antibody
- Suitable for use in mouse, rat, and human samples
- Can be used for western blotting and fluorescent imaging
