Apoptosis Assays for Advanced High-Throughput Flow Cytometry
Apoptosis is an essential process for normal tissue development and homeostasis by which cells undergo timely programmed cell death. Aberrations in apoptotic signaling are implicated in a range of human pathologies including cancer, autoimmune disease and neurodegeneration. Induction of apoptosis leads, in most cases, to the depolarization of mitochondria, the activation of caspases and plasma membrane alterations. The ability to simultaneously measure a multitude of apoptotic markers allows confirmation of pathway and insight into mechanism of action over time.
Assay Concept
Introducing Apoptosis Assays for Flow Cytometry
Figure 1. Apoptotic markers measured by the iQue® Human 4-Plex Apoptosis Kit. 1) Mitochondrial depolarization, 2) Caspase 3/7, 3) Annexin V and 4) Non-viable. The kit components can be utilized either separately or combination in a no wash format. Apoptosis reagents can also be multiplexed with other iQue® reagents in no wash assays.
Key Advantages
Key Advantages of Apoptosis Assays
- Detect and confirm apoptosis through different pathways using the mix and match reagents
- Gain a robust determination of apoptosis progression and insight into mechanism of action
- Simple mix-and-read 96 and 384-well protocol with no washing and no fixing
Detect and Confirm Apoptosis Through Different Pathways Using the Mix and Match Reagents
Figure 2. Cytotoxic and cytostatic compounds produce different cell health profiles. Jurkats (1e6/mL) treated with a range of concentrations of 4 compounds for 24 hours. Before analysis with iQue®️ Human 4-Plex Apoptosis Kit.
(A) Caspase 3/7 expressing cells are shown in a heatmap from iQue Forecyt®️.
(B,C,D,E) Depolarized mitochondria, Caspase 3/7, Annexin V and Non-viable (respectively) cell percentages are shown.
Gain a Robust Determination of Apoptosis Progression and Insight Into Mechanism of Action
Figure 3. Cell health markers show a time dependent increase when treated with an apoptosis inducing drug.
Jurkats (1e6/mL) treated with a range of concentrations of Staurosporin. 20 µL samples were taken and analyzed after 2, 6 and 24 hours of treatment with 3 apoptosis reagents. (A) Heat map of Caspase positive cell (%) from 2 hours, (B) Mitochondrial depolarization, (C) Caspase and (D) Annexin V
Simple Mix-and-Read 96 and 384-Well Protocol With No Washing and No Fixing
Figure 4. Schematic of the simple, mix and read apoptosis assay.
Ordering Information
Ordering Information
| iQue®️ Human 4-Plex Apoptosis Kit | ||
|---|---|---|
Platform: Compatible with iQue®️ 3 BR/VBR & iQue®️ Plus BR/VBR | ||
| Available Sizes | Catalog Numbers | |
1 x 384 | ||
5 x 384 | ||
20 x 384 | ||
50 x 384 | ||
Individual Apoptosis Reagents | ||||
|---|---|---|---|---|
| Platform: Compatible with iQue®️ 3 BR/VBR & iQue®️ Plus BR/VBR | ||||
| Product | Detection Channel | Available Size | Catalog Number | |
| iQue® Human Caspase 3/7 | B/Green | 5 x 384 | ||
| iQue® Human Annexin V | B/Yellow | 5 x 384 20 x 384 50 x 384 | ||
| iQue® Mitochondrial Membrane Potential | R/Red | 5 x 384 | ||
| iQue® Cell Membrane Integrity | B/Red | 5 x 384 | ||
