 |
| 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 |
$275 |
|
| * Please call 888 7 ASSAYS
(888-727-7297) or email info@celltechnology.com
for volume pricing |
|
|
