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QIA115 Caspase-9 Detection Kit (FITC-LEHD-FMK)

Caspase-9 Detection Kit (FITC-LEHD-FMK) MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
QIA115
  
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Overview

Replacement Information

Key Spec Table

Detection Methods
Fluorescence
Description
Overview

This product has been discontinued.





A fast, convenient, and sensitive method to measure caspase-9 in living cells. The fluorescent marker, FITC-LEHD-FMK, is a labeled, cell-permeable, non-toxic inhibitor that binds irreversibly to activated caspase-9 in living cells. Caspase-9 can be detected by the measurement of fluorescence intensity in a fluorescent microplate reader, flow cytometer, or by fluorescence microscopy.
Catalogue NumberQIA115
Brand Family Calbiochem®
References
Product Information
Detection methodFluorescence
Form100 Tests
FormatFlow cytometry or fluorescence microscopy
Kit containsLabeled Caspase-9 Inhibitor (FITC-LEHD-FMK), Wash Buffer, Unlabeled Caspase Inhibitor (Z-VAD-FMK), and a user protocol.
Applications
Biological Information
Assay time1.5 h
Sample TypeIntact cells
Physicochemical Information
Emission max.
Excitation max.
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
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.
Protect from Light Protect from light
Avoid freeze/thaw Avoid freeze/thaw
Do not freeze Ok to freeze
Packaging Information
Transport Information
Supplemental Information
Kit containsLabeled Caspase-9 Inhibitor (FITC-LEHD-FMK), Wash Buffer, Unlabeled Caspase Inhibitor (Z-VAD-FMK), and a user protocol.
Specifications
Global Trade Item Number
Catalogue Number GTIN
QIA115 0

Documentation

Caspase-9 Detection Kit (FITC-LEHD-FMK) Certificates of Analysis

TitleLot Number
QIA115

Brochure

Title
Caspases and other Apoptosis Related Tools Brochure
User Protocol

Revision13-November-2008 JSW
Form100 Tests
FormatFlow cytometry or fluorescence microscopy
Detection methodFluorescence
Speciesa broad range of species
StorageUpon arrival store the entire contents of the kit at -20°C.
Principles of the assayThe activation of caspases plays a central role in apoptosis. This Calbiochem® brand Caspase-9 Detection Kit provides a fast, convenient, and sensitive method to measure caspase-9 in living cells. The fluorescent marker, FITC-LEHD-FMK, is a labeled, cell-permeable, non-toxic inhibitor that binds irreversibly to activated caspase-9 in living cells. Caspase-9 can then be detected by the measurement of fluorescence intensity (excitation max.: ~485 nm; emission max.: ~535 nm) in a fluorescent plate reader, by flow cytometry, or by fluorescence microscopy.
Materials provided• FITC-LEHD-FMK (Kit Component No. JA7626-100UL): 1 vial, 100 µl
• Wash Buffer (Kit Component No. JA7627-100ML): 2 bottle, 100 ml
• Z-VAD-FMK (Kit Component No. JA7628-10UL): 1 vial, 10 µl
Detailed protocolCaspase-9 Assay Procedures

Staining

1. Induce apoptosis in cells (1 x 106/ml) as desired. Concurrently incubate a control culture without induction. An additional negative control can be prepared by adding the caspase 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-LEHD-FMK to each tube and incubate for 0.5-1 h in a 37°C incubator with 5% CO2.
4. Centrifuge cells at 3,000 rpm for 5 min and remove supernatant.
5. Resuspend cells in 0.5 ml of wash buffer, and centrifuge again.
6. Repeat Step 5.

Proceed depending on method of analysis.

Quantification by Flow Cytometry

1. For flow cytometric analysis, resuspend cells in 300 µl of wash buffer and place on ice.
2. Analyze samples by flow cytometry using the FL-1 channel.


Detection by Fluorescence Microscopy

1. Resuspend cells in 100 µl of wash buffer.
2. Add one drop of the cell suspension to a microslide and cover with a coverslip.
3. Observe cells under a fluorescence microscope using a FITC filter. Caspase-9 positive cells appear to have bright green signals, whereas caspase-9 negative control cells show a much weaker signal.

Analysis by Fluorescence Plate Reader

1. Resuspend cells in 100 µl of wash buffer.
2. Transfer the cell suspension to each well of the black plate.
3. Measure the fluorescence intensity at Ex. 485 nm and Em. 535 nm. For 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.