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aCella™ - TOX
Bioluminescence Non Radioactive Cytotoxicity
- 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
It is usually desirable to use smaller numbers of TCells than are
needed for the 51 Cr – 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 Cr 51
assays. Sample Protocol published.
- HTS - Adaptable for High Throughput format
- Non-destructive assay allows monitoring of additional
|Introduction to aCella-TOX:
Cell Technology introduces aCella-TOX
, a new and
highly sensitive assay using our patented Coupled Luminescent
technology for the detection of cytotoxicity
. This assay quantitatively measures the
release of Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) from
Primary Cells, mammalian cell lines, bacterial cells(1,2,3)
Other enzyme release assays(5,6,7)
for example the Lactate Dehydrogenase (LDH) release assay
(5,6,7), are inconvenient and/or
slow and may suffer from low sensitivity as a result of the poor
signal and interference by serum or phenol red present in the media.
The ATP-release assay
(8) is inconvenient and much less
sensitive than aCella-TOX, and is unsuitable for
use in a cytotoxicity assay because the lytic signal is indirect.
aCella-TOX can work in both these media
formulations and allows overnight assays while retaining its
sensitivity. The sensitivity of aCella-TOX is also
greatly enhanced by the coupled luminescent signal-amplification
system, which yields a strong luminescent signal from even small
amounts of released enzyme.
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
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,
- cellular cytotoxicity (T cells)
- complement (2,3), 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 (2), which is impossible with any other reported
liquid-phase method. Please consult with us if you have an
application requiring specialized techniques.
|Use of aCella-TOX for Measurement of
Cell-Mediated (T Cells, ADCC, NK) or Complement-Mediated Cytolysis
|The following kits are