Application

Apoptosis Assays for Advanced High Throughput Flow Cytometry

Simultaneous Measurement of Apoptotic Markers

Apoptosis Pathway

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.
 
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Assay Concept

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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

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Detect and confirm apoptosis through different pathways using the mix and match reagents.

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Gain a robust determination of apoptosis progression and insight into mechanism of action.

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Simple mix-and-read 96 and 384-well protocol with no washing and no fixing.

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Detect and confirm apoptosis through different pathways using the mix and match reagents.

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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

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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

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Figure 4. Schematic of the simple, mix and read apoptosis assay.

Ordering Information

 

iQue®️ Human 4-Plex Apoptosis Kit
Platform: Compatible with iQue®️ 3 BR/VBR & iQue®️ Plus BR/VBR
Available SizesCatalog Numbers
1 x 38490053
5 x 38490054
20 x 38490155
50 x 38490156

 

Individual apoptosis reagents
Platform: Compatible with iQue®️ 3 BR/VBR & iQue®️ Plus BR/VBR
ProductDetection ChannelAvailable SizeCatalog Number
iQue® Human Caspase 3/7
B/Green
5 x 38491034
20 x 38491035
50 x 38491036
iQue® Human Annexin V
B/Yellow
5 x 38491030
20 x 38491031
50 x 38491032
iQue® Mitochondrial Membrane Potential
R/Red
5 x 38491038
20 x 38491039
50 x 38491040
iQue® Cell Membrane Integrity
B/Red
5 x 38490346
20 x 38490347
50 x 38490348