CaspGLOWTM Fluorescein Active Caspase Staining Kit

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Antigen
  • CG5370
  • DCP-1
  • DCP1
  • Dcp1
  • Dmel\\CG5370
  • ccp1
  • dcp-1
  • dcp1
  • l(2)01862
  • l(2)02132
  • Death caspase-1
  • Dcp-1
Reactivity
Mammalian
2
1
Application
Detection (D), Fluorescence Microscopy (FM), Flow Cytometry (FACS)
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Purpose The CaspGLOW™ Fluorescein Active Caspase Staining Kit provides a convenient and sensitive means for detecting activated caspases in living cells. The assay utilizes the caspase family inhibitor VAD-FMK conjugated to FITC (FITC-VAD-FMK) as a marker. FITC-VAD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspases in apoptotic cells.
Brand CaspGLOW™
Sample Type Cell Samples
Detection Method Fluorometric
Specificity The CaspGLOW™ Fluorescein Active Caspase Staining Kit provides a convenient and sensitive means for detecting activated caspases in living cells. The assay utilizes the caspase family inhibitor VAD-FMK conjugated to FITC (FITC-VAD-FMK) as a marker. FITC-VAD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspases in apoptotic cells. The FITC label allows for direct detection of the activated caspases in apoptotic cells by fluorescence microscopy, flow cytometry, or fluorescence plate reader.
Characteristics CaspGLOWTM Fluorescein Active Caspase Staining Kit: Convenient & Sensitive Kit to Detect Activated Caspases in Living Cells. Detection Method: Fluorescence Microscopy, Flow Cytometry or Fluorescence Plate Reader.
Components FITC-VAD-FMK
Wash Buffer
Z-VAD-FMK
Alternative Name Caspase (CASP ELISA Kit Abstract)
Background Activation of caspases plays a central role in apoptosis. The CaspGLOW™ Fluorescein Active Caspase Staining Kit provides a convenient and sensitive means for detecting activated caspases in living cells. The assay utilizes the caspase family inhibitor VAD-FMK conjugated to FITC (FITC-VAD-FMK) as a marker. FITC-VAD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspases in apoptotic cells.
Application Notes Sensitive detection of activated caspases in living cells.
Comment

Further details regarding sample type: Live cells

Protocol A. Staining Procedure:
1. Induce apoptosis in cells (1 x 10^6 /mL) by desired method. Concurrently incubate a control culture without induction. An additional control can be prepared by adding the caspase family inhibitor Z-VAD-FMK at 1 µL/mL to an induced culture to inhibit caspase activation.
2. Aliquot 300 µL each of the induced and control cultures into eppendorf tubes.
3. Add 1 µL of FITC-VAD-FMK into each tube and incubate for 0.5-1 hour at 37 °C incubator with 5 % CO 2.
4. Centrifuge cells at 3000 rpm for 5 minutes and remove supernatant.
5. Resuspend cells in 0.5 mL of Wash Buffer, and centrifuge again.
6. Repeat Step
5. Proceed to B, C, or D depending on methods of analysis. B. Quantification by Flow Cytometry: For flow cytometric analysis, resuspend cells in 300 µL of Wash buffer. Put samples on ice. Analyzing samples by flow cytometry using the FL-1 channel. C. Detection by Fluorescence Microscopy: For fluorescence microscopic analysis, resuspend cells in 100 µL Wash buffer. Put one drop of the cell suspension onto a microslide and cover with a coverslip. Observe cells under a fluorescence microscope using FITC filter. Caspase positive cells appear to have brighter green signals, whereas caspase negative control cells show much weaker signal. D. Analysis by Fluorescence Plate Reader: For analysis with fluorescence plate reader, resuspend cells in 100 µL 1X Wash Buffer and then transfer the cell suspension to each well in a black microtiter plate. Measure the fluorescence intensity at Ex. = 485 nm and Em. = 535 nm. For control, use wells containing unlabeled cells.
Restrictions For Research Use only
Storage -20 °C
Expiry Date 6-12 months
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