Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit Green Fluorescence

Ordering information

Price
Catalog Number
Unit Size
Quantity

Add to cart

Additional ordering information

Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
International	See distributors
Bulk request	Inquire
Custom size	Inquire
Shipping	Standard overnight for United States, inquire for international

Spectral properties

Extinction coefficient (cm ^-1 M ^-1)	80000
Excitation (nm)	500
Emission (nm)	522

Storage, safety and handling

H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
UNSPSC	12352200

Alternative formats

Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit *Blue Fluorescence*

Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit *Red Fluorescence*

Overview SDS Protocol

Platform

Fluorescence microplate reader

Excitation	490 nm
Emission	525 nm
Cutoff	515 nm
Recommended plate	Black wall/clear bottom
Instrument specification(s)	Top/Bottom read mode

Components

Example protocol

AT A GLANCE

Protocol summary

Prepare cells with test compounds
Add equal volume of Caspase 9 Substrate working solution
Incubate at room temperature for 30 min to 1 hour
Monitor the fluorescence at Ex/Em = 490/525 nm

Important notes
Thaw all the kit components to room temperature before use.

PREPARATION OF WORKING SOLUTION

Add 50 μL of Caspase 9 Substrate (Component A) into 10 mL of Assay Buffer (Component B) and mix them well. Note: Aliquot and store the unused Caspase 9 Substrate (Component A) and Assay Buffer (Component B) at -20 ^oC. Avoid repeated freeze/thaw cycles.

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

SAMPLE EXPERIMENTAL PROTOCOL

Treat cells by adding 10 µL/well of 10X test compounds (96-well plate) or 5 µL/well of 5X test compounds (384-well plate) into PBS or the desired buffer. For blank wells (medium without the cells), add the same amount of compound buffer.
Incubate the cell plate in a 5% CO₂, 37°C incubator for a desired period of time to induce apoptosis.
Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) of Caspase 9 Substrate working solution.
Incubate the working solution plate at room temperature for at least 1 hour, protected from light. Note: If desired, add 1 µL of the 1 mM Ac-LEHD-CHO caspase 9 inhibitor to selected samples 10 minutes before adding the assay working solution at room temperature to confirm the inhibition of the caspase 9-like activity.
Centrifuge the cell plate (especially for the non-adherent cells) at 800 rpm for 2 minutes (brake off).
Monitor the fluorescence intensity at Ex/Em = 490/525 nm (Cutoff=515 nm).

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Extinction coefficient (cm ^-1 M ^-1)	80000
Excitation (nm)	500
Emission (nm)	522

Product Family

Name	Excitation (nm)	Emission (nm)
Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit Blue Fluorescence	341	441
Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit Red Fluorescence	532	619

Images

Figure 1. Detection of caspase 9 activities using Cell Meter™ Caspase 9 Activity Apoptosis Assay Kit in Jurkat cells. Jurkat cells were seeded at 200,000 cells/90 µL/well in a Costar black wall/clear bottom 96-well plate. The cells were treated with 10 mM adenosine for 48 hours while the untreated cells were used as control. The caspase 9 Substrate working solution (100 µL/well) was added and incubated at room temperature for 1 hour. The fluorescence intensity was measured at Ex/Em = 490/525 nm with a FlexStation™ microplate reader (Molecular Devices).

Citations

View all 3 citations: Citation Explorer

Contact-dependent signaling triggers tumor-like proliferation of CCM3 knockout endothelial cells in co-culture with wild-type cells
Authors: Rath, Matthias and Schwefel, Konrad and Malinverno, Matteo and Skowronek, Dariush and Leopoldi, Alexandra and Pilz, Robin A and Biedenweg, Doreen and Bekeschus, Sander and Penninger, Josef M and Dejana, Elisabetta and others,
Journal: Cellular and Molecular Life Sciences (2022): 1--20

Combination of Goniothalamin and Sol-Gel-Derived Bioactive Glass 45S5 Enhances Growth Inhibitory Activity via Apoptosis Induction and Cell Cycle Arrest in Breast Cancer Cells MCF-7
Authors: Bakar, Siti Aishah Abu and Ali, Abdul Manaf and Noor, Siti Noor Fazliah Mohd and Hamid, Shahrul Bariyah Sahul and Azhar, Nur Asna and Mohamad, Noor Muzamil and Ahmad, Nor Hazwani
Journal: BioMed Research International (2022)

The BH3 mimetic ($\pm$) gossypol induces ROS-independent apoptosis and mitochondrial dysfunction in human A375 melanoma cells in vitro
Authors: Haasler, Lisa and Kondadi, Arun Kumar and Tsigaras, Thanos and von Montfort, Claudia and Graf, Peter and Stahl, Wilhelm and Brenneisen, Peter
Journal: Archives of Toxicology (2021): 1349--1365

References

View all 31 references: Citation Explorer

Discovery of substituted N'-(2-oxoindolin-3-ylidene)benzohydrazides as new apoptosis inducers using a cell- and caspase-based HTS assay
Authors: Sirisoma N, Pervin A, Drewe J, Tseng B, Cai SX.
Journal: Bioorg Med Chem Lett (2009): 2710

Discovery of 1-benzoyl-3-cyanopyrrolo[1,2-a]quinolines as a new series of apoptosis inducers using a cell- and caspase-based high-throughput screening assay. 2: Structure-activity relationships of the 4-, 5-, 6-, 7- and 8-positions
Authors: Kemnitzer W, Kuemmerle J, Jiang S, Sirisoma N, Kasibhatla S, Crogan-Grundy C, Tseng B, Drewe J, Cai SX.
Journal: Bioorg Med Chem Lett (2009): 3481

Selective identification of cooperatively binding fragments in a high-throughput ligation assay enables development of a picomolar caspase-3 inhibitor
Authors: Schmidt MF, El-Dahshan A, Keller S, Rademann J.
Journal: Angew Chem Int Ed Engl (2009): 6346

Modified caspase-3 assay indicates correlation of caspase-3 activity with immunity of nonhuman primates to Yersinia pestis infection
Authors: Welkos S, Norris S, Adamovicz J.
Journal: Clin Vaccine Immunol (2008): 1134

Liver apoptosis following normothermic ischemia-reperfusion: in vivo evaluation of caspase activity by FLIVO assay in rats
Authors: Cursio R, Colosetti P, Auberger P, Gugenheim J.
Journal: Transplant Proc (2008): 2038

Discovery of 4-aryl-4H-chromenes as a new series of apoptosis inducers using a cell- and caspase-based high throughput screening assay. 4. Structure-activity relationships of N-alkyl substituted pyrrole fused at the 7,8-positions
Authors: Kemnitzer W, Drewe J, Jiang S, Zhang H, Crogan-Grundy C, Labreque D, Bubenick M, Attardo G, Denis R, Lamothe S, Gourdeau H, Tseng B, Kasibhatla S, Cai SX.
Journal: J Med Chem (2008): 417

Discovery of (naphthalen-4-yl)(phenyl)methanones and N-methyl-N-phenylnaphthalen-1-amines as new apoptosis inducers using a cell- and caspase-based HTS assay
Authors: Jiang S, Crogan-Grundy C, Drewe J, Tseng B, Cai SX.
Journal: Bioorg Med Chem Lett (2008): 5725

A dual-step fluorescence resonance energy transfer-based quenching assay for screening of caspase-3 inhibitors
Authors: Valanne A, Malmi P, Appelblom H, Niemela P, Soukka T.
Journal: Anal Biochem (2008): 71

Discovery of 5-(4-hydroxy-6-methyl-2-oxo-2H-pyran-3-yl)-7-phenyl-(E)-2,3,6,7-tetrahydro -1,4-thiazepines as a new series of apoptosis inducers using a cell- and caspase-based HTS assay
Authors: Drewe J, Kasibhatla S, Tseng B, Shelton E, Sper and io D, Yee RM, Litvak J, Sendzik M, Spencer JR, Cai SX.
Journal: Bioorg Med Chem Lett (2007): 4987

Split GFP complementation assay: a novel approach to quantitatively measure aggregation of tau in situ: effects of GSK3beta activation and caspase 3 cleavage
Authors: Chun W, Waldo GS, Johnson GV.
Journal: J Neurochem (2007): 2529