 |
| Caspase
/ Apoptosis
Detection |
 |
High
Throughput Screening
- Apo3HTS |
|
Caspase
Poly,1,2,3,7,8,9,10 Detection - FAM-FMK |
|
Caspase
Poly, 1,2,3,8,9,10, Detection
- (SR-FMK) |
|
Antibody
Specific Caspase 3 Detection
- Apo Active
3 (FITC) |
|
Antibody
Specific Caspase 3 Detection
- Apo Active
3 (PE) |
|
| NOS/ROS
Detection Kits |
|
| Fluorescent
Enzymatic Assays |
|
| Mitochondria
Membrane Potential detection |
|
| aCella
- Bioluminescence Assays Kits |
|
| Cytotoxicity |
|
| ELISA
Products |
|
| Cathepsin
B,K,L Detection Kits |
|
|
|
|
| |
ACT
1™
Assay for CytoToxicity
A non-radioactive cytotoxicity assay for flow cytometry |
| Key Benefits: |
-
No radioactive materials required
-
Applications - Works with a flow
cytometer or fluorescence microscope
-
Detects cytolytic
activity at a cellular level
-
Works with multiple
types of mammalian cell lines
|
| Technology: |
The most commonly
used method to measure CMC/ADCC is a radioactive chrominum-51
(51Cr) release assays (2). There are several disadvantages
with this assay: it is expensive, difficult to load
certain cell types, expensive to dispose of due to
strict environmental regulations, and has high background
from spontaneous release of 51Cr. With the use of flow
cytometry, it is now possible to eliminate the need
for radioactive material and increase the ability to
quantify cytolytic activity on a single cell bases.
Various groups have demonstrated that measuring CMC/ADCC
activity by flow cytometry has a strong (95%) correlation
with the traditional 51Cr release assay (3,4,5,6). |
| Assay Principle : |
A
cell tracking dye CFSE ( 7,8,9) is utilized to label
the target cell population. After the assay has run
its experimental protocol, 7AAD (live/dead) (10,11)
is added to measure cell death. 7AAD only enters membrane-compromised
cells and binds to DNA.
Flow cytometry is utilized to gate on the target
cells and measure 7AAD negative vs 7AAD postitve
cells. % cytotoxicity is calculated by the following
equation (see experimental example below):
7AAD positive (upper right quadrant)=R1/ 7AAD Postitve
(upper right quadrant)= R1 + 7AAD negative (lower
right quadrant)=R2 x100
|
| Citations |
|
| References |
-
a. Perussia,
B., (1998). Current Topics in Microbiology and
Immunology 230, p63.
b. Whiteside, T.L., Rinaldo, C.R. and Herberman,
R.B. (1992) Cytolytic Cellfunctions. In: N.R. Rose,
E.C. de Macario (Eds.), Manual of Clinical Laboratory
Immunology. American Society for Microbiology.
Washington, DC, p. 220.
-
Brunner, K.T., Manuel, J., Cerotini,
J.C., Chapuis, B., (1968). Quantitative Assay of
the
Lytic Action of Immune Lymphoid Cells on Cr-labelled
Allogenic Target Cells in- vitro; Inhibition
by Iso-antibody and by Drugs, Immunology 14,181.
-
Lee-MacAry, A.E., Ross, E.L, Davies,
D., and Wilkinson, R.W., (2001). Development of a
Novel
Flow Cytometric Cell-mediated Cytotoxcity Assay
Using the Fluorophores PKH-26 and TO-PRO-3
Iodide. J. Immunology. Met 252, 83-92.
-
Gogoy-Ramirez, K., Franck, K.,
and Gains, H., (2000). A Novel Method for the Simultaneous
Assessment
of Natural Killer Cell Conjugate Formation
and Cytotoxicity at the Single-cell Level by Multi-parameter
Flow Cytometry. Journal of Immunology. Met
239,
35-44.
-
Goldberg, J.E., Sherwood, S.W.,
Clayberger, C., (1999). A Novel Method for Measuring
CTL and
NK Cell-mediated Cytotoxicity Using Annexin
V and Two-color Flow Cytometry. Journal of Immunology.
Methods 224, 1.
-
Hatam, L,. Schuval, S., Bonagura,
V.R., (1994). Flow Cytometric Analysis of Natural
Killer Cell
Function as a Clinical Assay. Cytometry 16,59.
-
L.S De Clerck et al.,J. Immunol.
Meth. 172, 115 (1994).
-
M. Bronner-Fraser,J. Cell Biol.
101, 610 (1985).
-
M. Bronner-Fraser,J. Cell Biol.
101, 610 (1985).
-
Rabinovitch, P.S., et al., J.
Immunol. 136, 2769 (1986).
-
Su, X.,J. Immunol. 156, 156,
4198 (1996).
-
Olin, MR. Lee, J. Choi, K, and
Molitor, Tw. gd T-lymphocyte Cytotoxic Activity against
Mycobaterium
bovis Analysed by Flow Cytometry: Journal of
Immunological Methods; Publication in process.
-
Olin, MR. Thesis, K. Cho, J. and
Molitor, T.W. Morphine Suppresses Microglial Directed
Cytolytic
Activity by gd Lymphocytes. Journal of Neuroimmunology;
Publication in process.
|
| Kit contents: |
- 4 vials of CFSE membrane stain, part
no 4002.
- 3 vials of 7AAD Live/Dead stain, part no 4003.
|
| The following
kit is available: |
Product
|
Catalog No.
|
Size
(No. of Tests) |
Price |
ACT 1
Assay for CytoToxicity
|
ACT100-2 |
100 |
$245 |
|
| * Please call 888 7 ASSAYS (888-727-7297) or email info@celltechnology.com for
volume pricing |
|
|
