Millipore Sigma Vibrant Logo
Attention: We have moved. Merck Millipore products are no longer available for purchase on MerckMillipore.com.Learn More

QIA93 Caspase-3 Detection Kit (Red-DEVD-FMK)

Caspase-3 Detection Kit (Red-DEVD-FMK) MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
QIA93
  
Purchase on Sigma-Aldrich

Overview

Replacement Information

Key Spec Table

Detection Methods
Fluorescence
Description
OverviewA convenient and sensitive method for the detection of activated caspase-3 in living cells. The fluorescent marker is DEVD-FMK conjugated to sulforhodamine. This cell-permeable, non-toxic inhibitor binds irreversibly to activated caspase-3 in apoptotic cells. The fluorescence intensity can be measured by fluorescence microscopy, fluorescence microplate reader, or flow cytometry.
Catalogue NumberQIA93
Brand Family Calbiochem®
References
Product Information
Detection methodFluorescence
Form100 Tests
FormatFlow cytometry or fluorescence microscopy
Kit containsLabeled Caspase-3 Inhibitor (Red-DEVD-FMK), Wash Buffer, Caspase-3 Inhibitor (Z-VAD-FMK), and a user protocol.
Quality LevelMQ100
Applications
Biological Information
Assay time1.5 h
Sample TypeCultured 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: 23-26-36

Do not breathe fumes.
In case of contact with eyes, rinse immediately with plenty of water and seek medical advice.
Wear suitable protective clothing.
Product Usage Statements
Storage and Shipping Information
Ship Code Dry Ice Only
Toxicity Multiple Toxicity Values, refer to MSDS
Storage -20°C
Avoid freeze/thaw Avoid freeze/thaw
Do not freeze Ok to freeze
Packaging Information
Transport Information
Supplemental Information
Kit containsLabeled Caspase-3 Inhibitor (Red-DEVD-FMK), Wash Buffer, Caspase-3 Inhibitor (Z-VAD-FMK), and a user protocol.
Specifications
Global Trade Item Number
Catalogue Number GTIN
QIA93 0

Documentation

Caspase-3 Detection Kit (Red-DEVD-FMK) SDS

Title

Safety Data Sheet (SDS) 

Caspase-3 Detection Kit (Red-DEVD-FMK) Certificates of Analysis

TitleLot Number
QIA93

Brochure

Title
Caspases and other Apoptosis Related Tools Brochure
Kit SourceBook - 2nd Edition EURO
User Protocol

Revision07-October-2010 RFH
Form100 Tests
FormatFlow cytometry or fluorescence microscopy
Detection methodFluorescence
Speciesa broad range of species
BackgroundThe activation of caspases plays a central role in apoptosis. The Caspase-3 Detection Kit provides a convenient and sensitive means for detecting activated caspase-3 in situ in living cells. The assay utilizes a caspase-3 inhibitor (DEVD-FMK) conjugated to sulfo-rhodamine as the fluorescent in situ marker. Red-DEVD-FMK is cell permeable, nontoxic, and irreversibly binds to activated caspase-3 in apoptotic cells. The red fluorescence label allows for direct detection of activated caspase-3 in apoptotic cells by fluorescence microscopy, flow cytometry, or fluorescence plate reader.
Materials provided• Red-DEVD-FMK (Kit Component No. JA5900): 1 vial, 100 µl
• Wash Buffer (Kit Component No. JA5901): 2 bottles, 100 ml
• Z-VAD-FMK (Kit Component No. JA5902): 1 vial, 10 µl
Detailed protocolA. Staining Procedure:

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-3 inhibitor Z-VAD-FMK at 1 µl/ml to an induced culture to inhibit caspase-3 activation.
2. Aliquot 300 µl each of the induced and control cultures into microfuge tubes.
3. Add 1 µl of Red-DEVD-FMK into each tube and incubate for 0.5-1 h in a 37°C incubator with 5% CO2.
4. Centrifuge cells at 3000 rpm for 5 min and remove supernatant.
5. Resuspend cells in 0.5 ml of Wash Buffer and centrifuge again.
6. Repeat step 5.
7. Proceed to B, C, or D, depending upon method of analysis.

B. Quantification by Flow Cytometry

For flow cytometric analysis, resuspend cells in 300 µl of Wash buffer. Put samples on ice. Analyze samples by flow cytometry using the FL-2 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 the fluorescence microscope using a rhodamine filter. Caspase positive cells appear to have brighter red signals, whereas caspase negative control cells show much weaker signal.

D. Analysis by Fluorescence Plate Reader

For analysis with a fluorescence plate reader, resuspend cells in 100 µl Wash Buffer and transfer the cell suspension to each well of the black plate. Measure the fluorescence intensity at Ex. = ~540 nm and Em. = ~570 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.