Millipore Sigma Vibrant Logo
Atención: Nos hemos mudado. Los productos Merck Millipore ya no pueden adquirirse en MerckMillipore.comMás información

QIA89 Glutathione Apoptosis Assay Kit

Glutathione Apoptosis Assay Kit: Ficha de datos de seguridad (MSDS o SDS), certificado de análisis y de calidad (CoA y CoQ), expedientes, folletos y otros documentos disponibles.
QIA89
  
Purchase on Sigma-Aldrich

Descripción

Replacement Information

Tabla espec. clave

Detection Methods
Fluorometric
Description
Overview

This product has been discontinued.





A fluorometric assay to detect the in vitro changes in glutathione (GSH) levels during apoptosis. Utilizes monochlorobimane (MCB), a dye that fluoresces blue when bound to GSH. The adduct formation is catalyzed by glutathione S-transferase.
Catalogue NumberQIA89
Brand Family Calbiochem®
Materials Required but Not Delivered•Fluorimeter or fluorescence plate reader that can detect fluorescence at an Excitation max = ~380 nm and Emission max = ~460 nm
•PBS
•Apoptosis inducer of choice
References
Product Information
Detection methodFluorometric
Form100 Tests
Format96-well plate or cuvette
Kit containsCell Lysis Buffer, Monochlorobimane Reagent, GST Reagent, and a user protocol.
Applications
Key Applications Fluorometric
Biological Information
Assay time1.5-2 h
Sample TypeCell lysate
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: 26-36

In case of contact with eyes, rinse immediately with plenty of water and seek medical advice.
Wear suitable protective clothing.
Product Usage Statements
Intended useThe Calbiochem® Glutathione Apoptosis Detection Kit is intended for the measurement of changes in the glutathione level in cell lysates.
Storage and Shipping Information
Ship Code Dry Ice Only
Toxicity Multiple Toxicity Values, refer to MSDS
Storage -20°C
Storage ConditionsUpon arrival store the Monochlorobimane Reagent and GST Reagent at -20°C and the Cell Lysis Buffer at 4°C.
Avoid freeze/thaw Avoid freeze/thaw
Do not freeze Ok to freeze
Packaging Information
Transport Information
Supplemental Information
Kit containsCell Lysis Buffer, Monochlorobimane Reagent, GST Reagent, and a user protocol.
Specifications
Global Trade ITEM Number
Número de referencia GTIN
QIA89 0

Documentation

Glutathione Apoptosis Assay Kit Certificados de análisis

CargoNúmero de lote
QIA89

Folleto

Cargo
Caspases and other Apoptosis Related Tools Brochure
Kit SourceBook - 2nd Edition EURO
Kits SourceBook - 2nd Edition GBP
Protocolo de usuario

Revision06-Febuary-2009 RFH
Form100 Tests
Format96-well plate or cuvette
Detection methodFluorometric
StorageUpon arrival store the Monochlorobimane Reagent and GST Reagent at -20°C and the Cell Lysis Buffer at 4°C.
Intended useThe Calbiochem® Glutathione Apoptosis Detection Kit is intended for the measurement of changes in the glutathione level in cell lysates.
BackgroundGlutathione (GSH) is a low-molecular weight thiol that plays a critical role in the cellular defense against oxidative and nitrosative stress in mammalian cells. Diminished glutathione levels have been observed in the early stages of apoptosis.
Principles of the assayThe Glutathione Apoptosis Detection Kit, Fluorometric utilizes monochlorobimane (MCB), a dye that forms an adduct exclusively with GSH. The unbound MCB is nearly nonfluorescent, whereas the dye fluoresces blue when bound to glutathione (Excitation max = ~380 nm; Emission max = ~460 nm). The reaction is catalyzed by glutathione-S-transferase. Thus, the changes in glutathione levels during apoptosis can be easily detected using a fluorometer or a 96-well fluorometric plate reader.
Materials providedThere are sufficient reagents to perform 100 tests.

  • Cell Lysis Buffer (Kit Component No. JA5400-25ML): 1 bottle, 25 ml
  • Monochlorobimane Reagent (Kit Component No. JA5401-200UL): 1 vial, 200 µl, 25 mM in DMSO
  • GST Reagent (Kit Component No. JA5402-200UL): 1 vial, 200 µl, 50 U/ml
  • GSH standard (Kit Component No. JA5403-1MG): 1 vial, 1 mg
Materials Required but not provided•Fluorimeter or fluorescence plate reader that can detect fluorescence at an Excitation max = ~380 nm and Emission max = ~460 nm
•PBS
•Apoptosis inducer of choice
Detailed protocolNote: the following protocol has been optimized using Jurkat cells. Other cell types in which glutathione levels drop druing apoptosis may also be used, however, the assay may need to be optimized for other cell types.

1. Induce apoptosis in cells by desired method. Concurrently incubate control cells that are uninduced.
2. Collect 1 x 106 cells by centrifugation at 700 x g for 5 min at 4°C.
3. Remove the supernatant and resuspend the cell pellet in 1 ml ice-cold PBS.
4. Transfer to a 1.5 microcentrifuge tube and centrifuge at 700 x g for 5 min at 4°C. Remove the supernatant and discard.
5. Resuspend the cell pellet in 100 µl ice-cold Cell Lysis Buffer.
6. Incubate for 10 min on ice. Centrifuge in a microcentrifuge at 14,000 x g for 10 min.
7. Transfer the supernatant to a clean tube and discard the pellet.
8. Add 2 µl Monochlorobimane Reagent and 2 µl GST Reagent to each tube. Prepare a negative control sample by adding 1 µl Monochlorobimane Reagent to 100 µl Cell Lysis Buffer.
9. Incubate all samples and controls at 37°C for 15-30 min.
10. Measure the fluorescence using a fluorimeter or fluorescence plate reader with an excitation wavelength of ~380 nm and an emission wavelength of ~460 nm.
Registered TrademarksCalbiochem® is a registered trademark of EMD Chemicals, Inc.
Interactive Pathways™ is a trademark of EMD Chemicals, Inc.