SensiDeath - Ultra Sensitive Human Cell Death Assay

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Ultra-sensitive PCR-based human cell death assay

SensiDeath is a novel sensitive kit for human cell death detection. Taking benefit from qPCR high sensitivity, SensiDeath provides a simple method to detect cell death at its earliest stages by quantifying genomic DNA fragments released in the cytoplasm. Use it to get reliable measurements of apoptosis across a range of cell populations.

Catalogue Numbers:

IDY-SD-300 (Primer mix (20X): 300µL, SensiDeath Lysis buffer (6X): 25 mL)

IDY-SD-1000 (Primer mix (20X): 1 mL, SensiDeath Lysis buffer (6X): 85 mL)

Sizes:

Primer mix (20X): 300µL, SensiDeath Lysis buffer (6X): 25 mL (IDY-SD-300)

Primer mix (20X): 1 mL, SensiDeath Lysis buffer (6X): 85 mL (IDY-SD-1000)

Fast & simple
As simple as a qPCR. Lyse your cells, collect the cytosolic fraction and get results in less than 2h!

Extended dynamic range
Get reliable data and make sure to never miss out subtle changes in cell death.

Scalable
Compatible with 96-well plate format and efficient on small cell populations, SensiDeath is a convenient method for high-throughput cytotoxicity assays.

Early detection
Based on extremely sensitive cytosolic detection of genomic DNA, SensiDeath detects cell death as soon as it is initiated and up to its late phase.

How does SensiDeath work?

SensiDeath is based on a patented technology relying on the quantitation of genomic DNA released in the cell cytosol, a key process initiated upon cell death and a valuable marker of apoptosis. By amplifying genomic DNA present in isolated cytosolic fractions, you can quickly compare relative levels of apoptotic cells across a wide range of samples.

Steps:

Lyse your sample using the special lysis buffer disrupting cell plasma membrane specifically.
Centrifuge your lysate to recover the cytosolic fraction.
Amplify specific sequences present in the genome by quantitative PCR using our optimized primers.
Assess cell death that is directly proportional to cytosolic enrichment in genomic DNA.

Applications of SensiDeath

Outputs: comparative measurements of cell death levels for cytotoxic or viability assays. SensiDeath can be used as a reliable method to distinguish changes in apoptosis*.
*Check out our Results section for detailed results.

Sample types: human cells (adherent & suspension)*, tumoroids
*Check out the FAQ section for a full list of validated cell lines.

Analysis: PCR-based techniques (quantitative PCR, digital droplet PCR).

Kit contents of SensiDeath

All of these kits include a pair of optimized primers designed to amplify a genomic repeat DNA element. They have been thoroughly selected and optimized to combine high sensitivity, specificity and reproducibility in cell death detection.

These kits also include a concentrated lysis buffer, developed and optimized to recover cytosolic fractions by disrupting the cell plasma membrane specifically while maintaining integrity of the nuclear membrane as well as that of other cellular organelles.

Small kit (>300 assays*)

Primer mix (20X): 300 µL
SensiDeath Lysis buffer (6X): 25 mL

**Extended kit (>1000 assays*)**

Primer mix (20X): 1 mL
SensiDeath Lysis buffer (6X): 85 mL

*Number of assays are given for 20 µL qPCR reactions. More assays can be carried out with each kit if working with smaller qPCR reaction volumes.

Storage: the 6X lysis buffer can be stored at 4°C for at least 1 year. Primers can be stored at 4°C for one month, or at -20°C for long-term storage.

Additional resources:

Protocol

Results

SensiDeath provides increased sensitivity in cell death detection compared to existing methods

SensiDeath cell death sensitivity MOLM14 — Caspase-Glo® 3/7 or SensiDeath kits were used to measure cell death in increasing quantities of MOLM14 cells treated with 1 µM Etoposide for 16 hours (N=3). SensiDeath provides a 4-fold increase in detected signal compared to Caspase-Glo® 3/7 in treated samples and shows a lower detection limit.

Extended dynamic range of SensiDeath compared to existing methods

Annexin-V or SensiDeath methods were used to measure cell death in MOLM14 cells treated with increasing concentrations of Etoposide for 16 hours. Compared to Annexin-V, with which a signal upper limit is reached from 2.5 µM Etoposide, SensiDeath provides an extended dynamic range with additional intermediate values available up to 10µM Etoposide.

Image credits: Equipe ANUBIS, Université de Toulouse, France

Benchmarking of SensiDeath against other common methods

OCI-AML3 cells were treated for 16h with different doses of Aracytine and apoptosis was measured by different techniques on the equivalent of the indicated number of cells.

Image credits: Equipe ANUBIS, Université de Toulouse, France