Annexin V-FITC/PI Apoptosis Detection Kit

Apoptosis

Apoptosis refers to a highly regulated form of programmed cell death controlled by genes to maintain internal environmental stability. It is widely present in processes such as elimination of development, tissue homeostasis, infection or injury responses. Unlike cell necrosis, apoptosis is not a passive process, but an active process that involves the activation, expression, and regulation of a series of genes. It is an autonomous death process that exists to better adapt to the living environment. During the process of cell apoptosis, phenomena such as nuclear condensation, DNA breakage, cell wrinkling, plasma membrane vesicle formation, and the production of apoptotic bodies can occur, ultimately leading to cell death.

The dysregulation of cell apoptosis occurs in various disease states, including autoimmune diseases, neurodegenerative diseases, and cancer, and contributes to the pathological states of these diseases.

Induction pathway of cell apoptosis

Apoptosis can be induced through three different pathways:

1. Targeting mitochondrial function, including mitochondrial pathways, cellular pathways, or intrinsic apoptotic pathways. Mitochondrial dysfunction, decreased membrane potential, and increased mitochondrial permeability can be caused by DNA damage, oxidative stress, hypoxia, and other factors. Activation of caspase can cleave proteins within cells, leading to morphological changes such as apoptosis and ultimately cell death.

2. By directly transducing signals through adaptor proteins, such as activating cell surface death receptors (such as Fas, TNF receptors) and other apoptotic external pathways.

3. Perforin/granzyme signaling pathway for cell apoptosis: Granzyme A (a type of trypsin) and granzyme B (a serine protease) can independently activate cell apoptosis when delivered to target cells together with perforin (PFN) (a pore forming protein).

Annexin V-FITC/PI Apoptosis Detection Kit 

There are currently multiple methods for detecting apoptotic cells, including Annexin V assay, Caspase assay, mitochondrial function and membrane potential assay, DNA fragmentation and morphology assay, oxidative stress assay, ion concentration, pH value, and reactive oxygen species (ROS) level assay. The selection of appropriate detection methods is greatly influenced by their sensitivity and specificity, depending on the purpose of use and experimental model.

The Annexin V-FITC/PI Apoptosis Detection Kit (GLPBIO; GK10037) is a rapid and efficient detection kit for cell apoptosis levels. The work of Annexin V-FITC/PI Apoptosis Detection Kit mainly relies on FITC labeled recombinant human Annexin V protein. Annexin V is a Ca2+- dependent phospholipid binding protein with a molecular weight of 35-36kD, widely distributed in the cytoplasm of eukaryotic cells and involved in intracellular signal transduction. In normal cells, phosphatidylserine (PS) is only distributed on the inner side of the lipid bilayer of the cell membrane; In the early stage of cell apoptosis, phosphatidylserine undergoes a reversal, and FITC labeled Annexin V can specifically bind with phosphatidylserine that has been flipped out of the membrane, thereby labeling apoptotic cells. For necrotic cells, due to the loss of membrane integrity, Annexin V-FITC can enter the cytoplasm and bind with phosphatidylserine located on the inner side of the cell membrane, resulting in green fluorescence of necrotic cells.

Propidium iodide (PI) is a phenanthrene fluorescent dye that can bind to DNA. It can stain necrotic cells or cells that have lost membrane integrity in the late stage of apoptosis, presenting red fluorescence, while living cells and early apoptotic cells exhibit propidium iodide rejection.

Apoptosis can be detected and analyzed using flow cytometry, fluorescence microscopy, or other fluorescence detection devices.

Application scope of Annexin V-FITC/PI Apoptosis Detection Kit

The use of fluorescently labeled membrane associated protein V to measure membrane phosphatidylserine externalization is widely used for detecting cell apoptosis and has broad applications in scientific research.

1. Apoptosis research: applicable to various cell types, including mammalian cells, plant cells (requiring preparation of protoplasts), etc., to study the effects of various factors on cell apoptosis.

2. Drug screening: used to screen and evaluate drugs with anti apoptotic or pro apoptotic effects, and to study the effects of drugs on cell fate.

3. Disease research: Help researchers understand the mechanisms and regulation of cell apoptosis in various diseases such as tumors, neurodegenerative diseases, etc.

4. Toxicological research: Evaluating the effects of various chemicals or environmental factors on cell apoptosis.

Product Features

1. Combined with high efficiency.

2. Quick and convenient, with just one simple staining procedure that can be completed in 10-20 minutes.

3. This kit can clearly distinguish early and late apoptotic cells from necrotic cells through Annexin V-FITC and PI staining.