Cell Technology's products offer reliable solutions for cellular analysis and immuno-assay development and will be a great addition to our already large portfolio of Cell Viability and ELISA reagents. All Cell Tech products will soon be available on the ICT website.
In this post we will highlight aCella-TOX, a top-seller from the Cell Viability product line.
GAPDH is an important enzyme in the glycolysis and gluconeogenesis pathways. This homotetrameric enzyme catalyzes the oxidative phosphorylation of D-glyceraldehyde-3-phosphate to 1,3-diphosphoglycerate in the presence of co-factor and inorganic phosphate.
In the aCella-TOX reaction scheme, the release of GAPDH is coupled to the activity of the enzyme 3-Phosphoglyceric Phosphokinase (PGK) to produce ATP. ATP is detected via the luciferase, luciferin Bioluminescence methodology. Further, aCella-TOX is a homogeneous cytotoxicity assay; alternatively, in dual mode, aCella-TOX can measure cytotoxicity and cell viability in the same plate. Culture supernatants can also be removed from the original plate and assayed in a different plate, allowing kinetics runs to be set up. The assay is non-destructive, allowing the monitoring of additional parameters such as gene expression.
The aCella-TOX method has been tested with many modes of cytolysis, including cellular cytotoxicity (T cells) complement, pore-forming agents, antibiotic-mediated lysis of bacteria, and detergent mediated and mechanical lysis. The method is highly general, since all known cells express copious amounts of GAPDH, and, unlike other enzymes, GAPDH is very readily released from the cytoplasm upon cell lysis. Using specially adapted formulations, the sensitivity of the method can be driven below 1 eukaryotic cell, which is impossible with any other reported liquid-phase method.
- Safe - Non Radioactive Enzyme release assay
- Versatile - Useful for measuring activity of T cells, Primary Cells, NK, complement and other lytic agents
- Assay can be run in serum supplemented media
- Homogenous - One-step, no wash assay; assay can be run in same plate as samples
- FAST - Results in 3-5 minutes. Chromium 51 or europium release for measurement are time consuming. The inherent sensitivity of luciferase detection is enhanced by the amplification effect of enzyme turnover, which products thousands, millions, or billions of high energy molecules for each molecule of the enzyme
- Highly sensitive - can detect fewer than 500 cells/well in the presence of serum or as few as 10 cells/well in serum-free or heat-killed media
- GAPDH - the fact that GAPDH is a natural component of cells, and does not need to be introduced into the cells in any manner, distinguishes this assay from all methods which require pre-labeling of cells, transfection, transformation, or other methods of introducing proteins or other molecules into the target cells in order to generate a signal in a later step
- Advantages for measurement of cell mediated or complement mediated cytolysis. It is usually desirable to use smaller numbers of T cells than are needed for the 51Cr-release assay, since excessive numbers of effector cells can increase the background signal. This is now possible due to the high sensitivity of aCella-Tox
- ADCC/CMC Assays - a non-radioactive alternative to 51Cr assays
- HTS - adaptable for high throughput format
- Non-destructive assay allows monitoring of additional parameters
We are excited to bring this product and all the Cell Tech products into our product line. If you have any technical questions or would like more information on aCella-TOX, please feel free to contact us.
Look for more exciting features and changes ahead.