aCella - TOX 1000 Tests | CLATOX100-4

aCella - TOX 1000 Tests | CLATOX100-4 | Cell Technology

Description

Key Benefits

• 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 produces 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 prelabelling 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 TCells 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. Please click here for a direct comparison between the aCella-TOX and (51Cr) Chromium Release Methods
• HTS – Adaptable for High Throughput format
• Non-destructive assay allows monitoring of additional parameters..

Additional information

Kit Size:500, 1000

Includes: No Plates, 5 Lumi Plates, 5 Lumi Plates + 5 Tissue Culture Plates

ASSAY PRINCIPLE

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 cofactor 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.