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aCella™ - TOX
Bioluminescence Non Radioactive Cytotoxicity
Assay (GAPDH) |
| 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.
- 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 parameters.
|
| 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
(1).
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.
|
| 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.
|
| Applications: |
|
The aCella-TOX method has been tested with
many modes of cytolysis, including;
- 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 |
| Kit Content: |
- Component 1: 4x Enzyme Assay
Reagent.............................Part
6001
- Component 2: 1x Enzyme Assay Diluent
..............................Part 3008
- Component 3: Glyeraldehyde 3-Phosphate
(G3P)................Part 6003
- Component 4: 50x Detection
Reagent..................................Part
6002
- Component 5: 5.5x Detection Assay
Diluent........................Part 3009
- Component 6: Lytic
Agent....................................................Part#
3035
|
|
The following kits are available: |
Catalog
|
Size |
Price |
CLATOX 100-3
|
500 |
€595 |
| CLATOX 100-4 |
1000 |
€1145 |
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