Oxidative Stress Induced Apoptosis Assay Services
In some pathophysiological states, oxidative stress will occur when the free radical products in cells exceed their own antioxidant capacity. Oxidative damage induced by reactive oxygen species plays an important role in many chronic diseases, such as diabetes and atherosclerosis. Apoptosis, also known as programmed cell death, refers to the natural cell death process of nucleated cells by starting their internal mechanism under certain conditions, mainly through the activation of endogenous DNA endonuclease. Studies have shown that high-dose ROS can induce cell necrosis, while low-dose ROS can cause apoptosis of various cells. Creative Bioarray provides a variety of detection services suitable for oxidative stress apoptotic cells, providing assistance for the research of cell oxidative damage.
Cell Morphology Observation
The most classical morphological standard for ROS is apoptosis. We observed the morphology of apoptotic cells by light microscope eosin staining or Gimsa staining and electron microscope.
Measurement standard: chromatin concentration, nuclear membrane wrinkling and rupture, apoptotic body formation and so on can be observed in apoptotic cells.
Phosphatidylserine Valgus Analysis Service
One of the early changes of apoptosis induced by ROS is the shift of phosphatidylserine from inside to outside the cell membrane.
Determination standard: We used fluorescein isothiocyanate to label membrane adhesion protein 5, combined with propidium iodide for double staining of apoptotic cells, which can distinguish apoptotic cells from necrotic cells.
Mitochondrial Membrane Potential Energy Assay Service
Free radicals produced by ROS stimulated cells can directly act on mitochondria and reduce mitochondrial membrane potential. When the mitochondrial membrane potential drops below the threshold, it will induce apoptosis.
Assay service: We measured the fluorescence intensity of rhl23 in cells to reflect the changes of mitochondrial intimal potential.
DNA Fragmentation Assay Service
The main biochemical feature of apoptosis induced by oxidative stress is that its chromatin is concentrated, and the chromatin DNA breaks at the junction between nucleosome units to form a large DNA fragment of 50-300 KB long or an oligonucleotide fragment of 180-200 BP integral multiple. Therefore, the DNA electrophoresis of healthy living cells is a band, while the DNA electrophoresis of apoptotic cells appears a characteristic ladder band. There were two kinds of agarose gel electrophoresis and polyacrylamide gel electrophoresis. This method is simple, qualitative and quantitative, but it can not show the morphological and structural characteristics of tissues and cells and the relationship between cells and surrounding tissues.
3-OH Terminal Assay Service
In ROS induced apoptosis, a large number of viscous 3-OH terminals are produced by double strand breaks or single strand breaks of chromosomal DNA.
Determination method: under the action of deoxyribonucleic acid terminal transferase, we labeled the derivatives formed by deoxyribonucleic acid and fluorescein, peroxidase, alkaline phosphatase or biotin to the 3-OH end of DNA, so as to detect apoptotic cells.
Caspase-3 Activity Assay Service
Measurement standard: in the early stage of apoptosis, caspase-3 is activated, and the activated form of Caspase-3 consists of two large subunits (17 × 103) and 2 small subunits (12 × 103) and cleave the corresponding cytoplasmic and nuclear substrates. In the late stage of apoptosis and dead cells, the activity of Caspase-3 decreased significantly.
Methods: Western blot, fluorescence spectrophotometer and flow cytometry were used to detect caspase-3.
Detection of apoptosis related genes induced by oxidative damage
| Gene |
Assay Methods |
| Bcl-2 gene assay |
We mainly provide Western blot hybridization and real-time fluorescence quantitative method to detect it. With the development of fluorescence quantitative polymerase chain reaction in recent years, quantitative polymerase chain reaction is undoubtedly faster and more accurate than the former. |
| p53 gene assay |
Immunohistochemistry and in situ hybridization are often used to detect. |
| Nuclear factor-kB (NF-kB) assay |
Immunochemical staining, polyacrylamide gel electrophoresis, in situ hybridization, etc. |
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All services and products are for scientific use only, not for medical use！
