QIA90 Caspase Detection Kit (FITC-VAD-FMK)

QIA90
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      Overview

      Replacement Information

      Key Specifications Table

      Detection Methods
      Fluorescence

      Pricing & Availability

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      QIA90-1KIT
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          Description
          OverviewA convenient and sensitive method for the detection of activated caspases in living cells. The fluorescent marker, FITC-VAD-FMK, is a cell-permeable, non-toxic inhibitor that binds irreversibly to activated caspases in apoptotic cells. The fluorescence intensity can be measured by fluorescence microscopy, fluorescence plate reader, or flow cytometry.
          Catalogue NumberQIA90
          Brand Family Calbiochem®
          References
          Product Information
          Detection method Fluorescence
          Form100 Tests
          FormatFlow Cytometry or Fluorimeter
          Kit containsLabeled Caspase Inhibitor (FITC-VAD-FMK), Wash Buffer, Caspase Inhibitor (Z-VAD-FMK), and a user protocol.
          Applications
          Biological Information
          Assay time1.5 h
          Sample TypeIntact cells
          Species Reactivity
          • A Broad Range Of Species
          Physicochemical Information
          Emission max.
          Excitation max.
          Dimensions
          Materials Information
          Toxicological Information
          Safety Information according to GHS
          Safety Information
          R PhraseR: 36/37/38

          Irritating to eyes, respiratory system and skin.
          S PhraseS: 23-26-38

          Do not breathe fumes.
          In case of contact with eyes, rinse immediately with plenty of water and seek medical advice.
          In case of insufficient ventilation, wear suitable respiratory equipment.
          Product Usage Statements
          Storage and Shipping Information
          Ship Code Dry Ice Only
          Toxicity Multiple Toxicity Values, refer to MSDS
          Storage -20°C
          Storage ConditionsUpon arrival store the entire contents of the kit at -20°C.
          Avoid freeze/thaw Avoid freeze/thaw
          Do not freeze Ok to freeze
          Packaging Information
          Transport Information
          Supplemental Information
          Kit containsLabeled Caspase Inhibitor (FITC-VAD-FMK), Wash Buffer, Caspase Inhibitor (Z-VAD-FMK), and a user protocol.
          Specifications

          Documentation

          Caspase Detection Kit (FITC-VAD-FMK) SDS

          Title

          Safety Data Sheet (SDS) 

          Caspase Detection Kit (FITC-VAD-FMK) Certificates of Analysis

          TitleLot Number
          QIA90

          Brochure

          Title
          The Active Siteル Volume 4, Number 1
          User Protocol

          Revision10-August-2007 RFH
          Form100 Tests
          FormatFlow Cytometry or Fluorimeter
          Detection method Fluorescence
          Speciesa broad range of species
          StorageUpon arrival store the entire contents of the kit at -20°C.
          Principles of the assayThe Calbiochem® Caspase Detection Kit (FITC-VAD-FMK) provides a convenient and sensitive means for detecting activated caspases in situ in living cells. The assay utilizes a caspase inhibitor (VAD-FMK) conjugated to FITC as the fluorescent in situ marker. FITC-VAD-FMK is cell permeable and nontoxic and irreversibly binds to activated caspases in apoptotic cells. The FITC label allows for direct detection of activated caspases in apoptotic cells by fluorescence microscopy, flow cytometry, or fluorescence plate reader.
          Materials provided• FITC-VAD-FMK (Kit Component No. JA5600): 1 vial, 100 µl each
          • Wash Buffer (Kit Component No. JA5601): 2 bottles 100 ml each
          • Z-VAD-FMK (Kit Component No. JA5602): 1 vial, 10 µl
          Detailed protocolA. Staining Protocol:
          1. Induce apoptosis in cells (1 x 106/ml) by desired method. Concurrently incubate a control culture without induction. An additional control can be prepared by adding the caspase inhibitor Z-VAD-FMK at 1 µl/ml culture to an induced culture to inhibit caspase activity.
          2. Aliquot 300 µl each of the induced and control cultures into microfuge tubes.
          3. Add 1 µl FITC-VAD-FMK into each tube and incubate for 0.5-1 h in a 37°C incubator with 5% CO2.
          4. Centrifuge the cells at 3000 rpm for 5 min and discard supernatant.
          5. Resuspend the cells in 0.5 ml Wash Buffer and centrifuge as above. Repeat for a total of 2 washes.
          6. Proceed to analysis using one of the methods below.

          B. Quantification by Flow Cytometry
          For flow cytometric analysis, resuspend the cells in 300 µl Wash Buffer. Put samples on ice. Analyze samples by flow cytometry using the FL-1 channel.

          C. Detection by Fluorescence Microscopy
          For fluorescence microscopy, resuspend the cells in 100 µl Wash Buffer. Put one drop of the cell suspension onto a slide and cover with a coverslip. Observe cells under the fluorescence microscope using a 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 a fluorescence plate reader, resuspend the cells in 100 µl Wash Buffer and transfer the cell suspension to designated wells of the black plate. Measure the fluorescence intensity at an Exitation of ~485 nm and an Emission of = ~535 nm. For a control, use wells containing unlabeled cells.
          Registered TrademarksCalbiochem® is a registered trademark of EMD Chemicals, Inc.
          Interactive Pathways™ is a trademark of EMD Chemicals, Inc.