VF 488 Caspase 3 Assay Kit for Live Cells
Based on 1 Customer Validation
MCE VF 488 Caspase 3 Assay Kit for Live Cells uses a fluorescent substrate with cell membrane permeability to detect Caspase 3 activity in live cells.
-
Storage :
4℃, 3 years. Protect from light.
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
Description & Advantages
Cysteine-dependent aspartate-directed proteases (Caspases) are a family of cysteine proteases that play important roles in apoptosis, necrosis, and inflammation.
Caspase 3, a cysteine protease also known as CPP32, Yama or apopain, belongs to the CED-3 subfamily. Caspase 3 can cleave procaspase 2, 6, 7, and 9, and can directly and specifically cleave many caspase substrates, including PARP, ICAD, gelsolin, and fodrin. In addition, Caspase 3 also plays a key role in the process of cell apoptosis, including chromatin condensation and DNA fragmentation.
MCE VF 488 Caspase 3 Assay Kit for Live Cells uses a fluorescent substrate with cell membrane permeability to detect Caspase 3 activity in live cells. The principle is that the substrate peptide DEVD coupled to a fluorogenic DNA dye is initially non-fluorescent, penetrates the plasma membrane and enters the cytoplasm. In apoptotic cells, caspase3 cleaves the substrate, releasing DNA dye, which migrates to the cell nucleus and stains DNA with bright green fluorescence, allowing real-time detection of apoptosis in living cells.
Features of MCE VF 488 Caspase 3 Assay Kit for Live Cells
1. Bifunctional: It can be used to detect caspase-3 activity and observe the morphological changes in the nucleus during apoptosis.
2. Compatibility: It can be assayed by fluorescence microscopy, flow cytometry or Fluorence microplate reader. After detection, the sample can be fixed with paraformaldehyde and subsequently a variety of immunostaining such as immunofluorescence and immunohistochemistry can be performed for further analysis.
3. Simple and Fast: The reagents can be added directly to the cell culture medium and the assay can be completed in 15-30 min.
Protocol
1. Cell treatment
Culture and treat cells according to the experimental design. Include an untreated control, an apoptosis-induced group, and a Caspase 3 inhibitor control when appropriate.
2. Inhibitor control (optional)
Add Inhibitor (2 mM Ac-DEVD-CHO, DMSO) to the inhibitor-control group before adding the fluorescent substrate to verify Caspase 3-specific fluorescence.
3. Live-cell staining
1. Add Substrate (0.2 mM VF 488 Caspase 3, DMSO) directly to the culture medium. A 1 : 200 dilution is recommended as a starting condition, i.e. 5 μL Substrate per 1 mL medium.
2. Incubate at 37°C for 15-30 min protected from light.
3. Analyze directly by live-cell fluorescence microscopy, flow cytometry, or fluorescence microplate reader. VF 488-positive fluorescence reflects intracellular Caspase 3 activity in living cells.
4. After detection, samples may be fixed with paraformaldehyde for subsequent immunofluorescence or immunohistochemistry if required.
Note: This is a live-cell assay. Protect samples and reagents from light during handling and detection.
Storage
4℃, 3 years.
Protect from light.
Components
| Components | HY-K1088-25T | HY-K1088-100T |
|---|---|---|
| Substrate (0.2 mM VF 488 Caspase 3, DMSO) | 125 μL | 500 μL |
| Inhibitor (2 mM Ac-DEVD-CHO, DMSO) | 20 μL | 100 μL |
Please enter the correct Lot Number
