Caspase 1 Activity Assay Kit
Based on 1 publication(s) in Google Scholar
MCE Caspase 1 Activity Assay Kit provides a simple and convenient way for detecting the activity of detection of Caspase 1 activity that based on the spectrophotometric.
-
Storage :
-20°C, 1 year
Keep away 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. Sequential activation of caspases plays an important role in cell apoptosis. Caspase-1 (Interleukin 1β converting enzyme (ICE)) is an interleukin-1 converting enzyme that proteolytically cleaves the precursor protein of the inflammatory cytokines interleukin 18 and interleukin 1β into active forms, regulates apoptosis by shearing Bcl-XL and plays a key role in regulating host responses to inflammatory stimuli through its action on various cytokines.
MCE Caspase 1 Activity Assay Kit provides a simple and convenient way for detecting the activity of detection of Caspase 1 activity that based on the spectrophotometric. Caspase 1 catalyzes the substrate Ac-YVAD-pNA and produces yellow pNA, and the luminescence of the pNA is quantified using a spectrophotometer at 405 nm to determine the activity of Caspase 1.
Protocol
1. Sample Preparation
1. Collect approximately 1 × 106-5 × 106 treated cells and wash once with PBS.
2. Resuspend cells in approximately 100 μL pre-cooled Lysis Buffer and lyse on ice for approximately 10-15 min.
3. Centrifuge at approximately 16,000 × g for 10-15 min at 4°C and collect the supernatant.
4. Determine protein concentration and normalize total protein among samples.
2. pNA Standard Curve
Prepare serial standards using the supplied pNA (10 mM) and measure absorbance at 405 nm to quantify released pNA.
3. Caspase 1 Activity Reaction
| Component | Per reaction |
|---|---|
| Protein lysate supernatant | 50 μL |
| Assay Buffer | 40 μL |
| Ac-YVAD-pNA (2 mM) | 10 μL |
1. Mix gently and incubate at 37°C for 1-2 h protected from light.
2. Measure absorbance at 405 nm.
3. Calculate Caspase 1 activity from the pNA standard curve or from the A405 increase relative to the control.
Note: Caspase 1 cleaves Ac-YVAD-pNA to release yellow pNA. Replicate wells are recommended, and equal total protein should be used for all samples.
Publications
-
Journal Impact Factor
-
Most Recent
Storage
-20°C, 1 year
Keep away from light
Components
| Components | HY-K2610-100T |
|---|---|
| Lysis Buffer | 20 mL |
| Assay Buffer | 10 mL |
| Ac-YVAD-pNA (2 mM) | 1 mL |
| pNA (10 mM) | 1 mL |
Please enter the correct Lot Number
