SensiDeath is a kit for the detection of cell death. Taking advantage of the high sensitivity of qPCR, SensiDeath provides a simple method to detect cell death at earlier stages by quantifying genomic DNA fragments released into the cytoplasm.
Return to previous page
SensiDeath
How does it work?
SensiDeath is based on the quantification of genomic DNA released into the cell cytosol, a key process initiated after cell death and a valuable marker of apoptosis. By amplifying genomic DNA present in isolated cytosolic fractions, relative levels of apoptotic cells in a wide range of human cell samples can be rapidly compared.
The protocol is very simple:
1- Lyse the cells using the buffer solution included in the kit, a solution that specifically alters the plasmatic cell membrane.
2- Centrifuge the lysate to recover the cytosolic fraction.
3- Amplify the specific sequences present in the genome by quantitative PCR using the optimized primers of the kit.
4- Evaluates cell death, directly proportional to incrment of genomic DNA in the cytosol.
-
-
-
- Sample types: human cells (adherent and in suspension)
- Analysis: PCR-based techniques (quantitative PCR, digital droplet PCR ).
-
-
Frequently Asked Questions:
What samples can be analyzed with SensiDeath?
SensiDeath has been successfully used to detect cell death in a variety of human cell lines, including:
– Adherent cells: HepG2, MDA-MB-231, HeLa, SK-Hep, HEK293, HUVEC, primary fibroblasts, etc.
– Suspension cells: OCI-AML3, MOLM14, MV4-11, HL-60, THP-1, U937, etc.
This list is not exhaustive and SensiDeath could be used to assess cell death in any other human cell type.
Do you require any specific material or equipment?
SensiDeath kits contain a concentrated lysis buffer and primers optimized for genomic DNA amplification. The only reagent needed is nuclease-free water and any SYBR Green qPCR
As for equipment, a simple benchtop centrifuge is needed for sample preparation and a thermocycler to perform qPCR. No sophisticated equipment is needed.
What phase(s) of apopotosi is SensiDeath detecting?
The presence of nuclear DNA in the cytosol can be detected in the early, middle and late phases of apoptosis. The cytosolic concentration of nuclear DNA will be highest in the middle phase, but the extreme sensitivity of the SensiDeath method allows it to be detected also in its earliest phases.
Can mitochondrial DNA influence the assay?
No, our lysis buffer supplied in the kit has been specifically optimized to rupture the plasma membrane of the cell, leaving the nuclear membrane, as well as that of all other organelles, perfectly intact. Therefore, there is no risk of recovering mitochondrial DNA in the isolated cytosolic fractions.
Can I use another lysis buffer than the one included in the kit?
The SensiDeath lysis buffer included in the kit has been specifically developed to destroy the plasma membrane of the cell while keeping the membrane of nuclei and other organelles intact, allowing specific recovery of the cytosolic fraction. Furthermore, its components have been optimized not to interfere with subsequent qPCR reactions. Therefore, we do not recommend the use of other lysis buffers, as we cannot guarantee the accuracy of the cell death measurements obtained.
I am text block. Click edit button to change this text. Lorem ipsum dolor sit amet, consectetur adipiscing elit. Ut elit tellus, luctus nec ullamcorper mattis, pulvinar dapibus leo.
If you would like more information about Idylle or this product, please do not hesitate to contact us:
