Advantage of Apoptosis-related ELISA Method

Apoptosis, also known as programmed cell death, is a natural process or physiological death of cells in the regulation of a series of endogenous genes. Apoptosis and necrosis are two different forms of cell death, and they can be distinguised by their characteristics in morphology, biochemistry and molecular biology.

There are several traditional methods of detecting apoptosis, including:

1. Apoptosis morphological examination, including observe the presence of apoptotic bodies by using optical microscope, inverted microscope, fluorescence microscope, CLSM or TEM;

2. Phosphatidylserine analysis (AnnexinV method), including using labeled Annexin-V as a fluorescent probe, detect the apoptosis state by flow cytometry or fluorescence microscopy;

3. DNA ladder detection, also known by TUNEL (terminal dexynucleotidyl transferase(TdT)-mediated dUTP nick end labeling), also known as end-labeling of DNA fragmentation in situ, this method can label the 3'-OH of DNA molecules gap in situ with an observable markers such as fluorescein, which can label nuclear DNA of apoptotic cells in situ, then the cells can be observed with a fluorescent microscope.

However, the above apoptosis detection methods are mostly qualitative rather than quantitative, so it is difficult to use those data to reflect the level of apoptosis, while flow cytometry can used to detect apoptosis quantitatively, it is still hard for beginners who lack enough experimental skills to get a stable result. With the continuous development of genetic engineering techniques, apoptosis-related ELISA  products are rising increasing attention in researchers.

Caspase family have a critical role in apoptosis-mediated processes, caspase-3 , as a key executive molecule, plays an important function in many apoptosis signaling pathways. Caspase-3 zymogen (32KD) is generally present in the cytoplasm, in the early stages of apoptosis, which is activated, the activation of caspase-3 is composed of two large subunits (17KD) and two small subunits (12KD), cleaved into corresponding cytoplasm nucleus substrate, ultimately leading to apoptosis. But in the late stage of apoptosis and dead cells, caspase-3 activity was significantly decreased. According to these characteristics, Cloud-Clone Corp. has developed a caspase 3 (CASP3, caspase-3) detection kit (SEA626Hu), which may be quantitative determination of CASP3 in human serum, plasma, tissue homogenates, cell lysate or other biological fluids.

In addition, Cloud-Clone Corp. has developed some of apoptosis regulatory protein assay kit, including but not limited to: apoptosis-related factor (FAS) assay kit (SEA030Hu), apoptosis-related factor ligand (FASL) detection kit (SEA031Hu), B- cell lymphoma factor 2 (Bcl2) detection kit (SEA778Hu), Bcl2 associated X protein (Bax) detection kit (SEB343Hu), tumor protein p53 (TP53) detection kit (SEA928Hu). On the other hand, in order to meet the needs of different experimenters, Cloud-Clone Corp. also developed apoptosis-related product for other specie, such as rats, mice, cattle and other species, these products provide very good tools (for the detailed information about apoptosis detection kit details, please see: apoptosis-related protein family detection products) test kits are to detect apoptosis.

Compared with other conventional methods to detect apoptosis, apoptosis-related ELISA assay is a relatvely simple, time-saving method with high sensitivity, this assay also can detect a large number of samples at one time, so we summarized the characteristics of those methods as follows :

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