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Caspase 3 and caspase 9 colorimetric assay kits

Manufactured by R&D Systems
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The Caspase-3 and Caspase-9 Colorimetric Assay Kits provide a simple and effective method for measuring the activities of caspase-3 and caspase-9, two key enzymes involved in the apoptosis (programmed cell death) pathway. The assay kits utilize colorimetric substrates that, upon cleavage by the target caspases, release a chromophore detectable by spectrophotometry. This allows for the quantitative analysis of caspase-3 and caspase-9 activity in cell and tissue samples.

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Lab products found in correlation

Penicillin, by Thermo Fisher Scientific (3 mentions) Streptomycin, by Thermo Fisher Scientific (3 mentions) Trypsin edta, by Thermo Fisher Scientific (3 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Muse caspase 3 9 assay kits, by Merck Group (1 mentions) Calpeptin, by Merck Group (1 mentions) Trypan blue, by Merck Group (1 mentions) Mitochondria cytosol fractionation kit, by Abcam (1 mentions) 1 2 bis 2 aminophenoxy ethane n n n n tetraacetic acid tetrakis acetoxymethyl ester bapta am, by Merck Group (1 mentions) 2 7 dichlorodihydrofluorescein diacetate h2dcfda, by Thermo Fisher Scientific (1 mentions) 3 3 dihexyloxacarbocyanine iodide, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin solution, by GE Healthcare (1 mentions) Thiazolyl blue tetrazolium bromide mtt, by Merck Group (1 mentions) Curcumin, by Merck Group (1 mentions) Growth supplement, by Thermo Fisher Scientific (1 mentions) Trypsin edta solution, by Thermo Fisher Scientific (1 mentions) Culture media, by Thermo Fisher Scientific (1 mentions) Anti mouse igg hrp antibody, by Santa Cruz Biotechnology (1 mentions) Anti bax antibody, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

Colorimetric assay kit (46 citations) Caspase-3 Colorimetric Assay kit (31 citations) Caspase-3 (21 citations) Caspase-3 assay kit (8 citations) Colorimetric assay (8 citations) Caspase Colorimetric Assay Kit (6 citations) Caspase-3 Colorimetric Assay (6 citations) Caspase-9 (6 citations) Caspase-3 colorimetric kit (2 citations)

9 protocols using caspase 3 and caspase 9 colorimetric assay kits

1

Caspase-3 and Caspase-9 Activity Assay

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CAR cells (2 × 105 cells/ per well) were seeded into 6-well plates and incubated with 0, 25, 50, 75 and 100 of YC-1 for 48 h. At the end of the treatment, cells were harvested and cell lysates were assessed in accordance with the manufacturer’s instruction provided in the caspase-3 and caspase-9 Colorimetric Assay kits (R&D Systems Inc.). Cell lysate protein was then incubated for 1 h at 37 °C with specific caspase-3 substrate (DEVD-pNA) or caspase-9 substrate (LEHD-pNA) in the reaction buffer (provided in the kits). The OD405 of the released pNA in each sample was measured as previously described [86 (link), 105 (link)].
Lee M.R., Lin C., Lu C.C., Kuo S.C., Tsao J.W., Juan Y.N., Chiu H.Y., Lee F.Y., Yang J.S, & Tsai F.J. (2017). YC-1 induces G0/G1 phase arrest and mitochondria-dependent apoptosis in cisplatin-resistant human oral cancer CAR cells. BioMedicine, 7(2), 12.
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2

Quercetin and Caspase Expression in Cells

Cited 1 time
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Cells (1 × 106 cells/mL) were cultured in a 10-cm dish with 50 or 75 μM quercetin and then treated with 500 μM indomethacin for 24 h for the measurement of caspase protein expression. Before being harvested through centrifugation (400× g; Caspase-3 and Caspase-9 Colorimetric Assay Kits, R&D Systems Inc., Minneapolis, MN, USA), the cells were cultured in the working solution from the Muse Caspase-3/9 Assay Kits (Millipore; Merck KGaA) in accordance with manufacturer protocols [95 (link),97 (link)].
Chen C., Yang J.S., Lu C.C., Wu Y.T, & Chen F.A. (2021). Effect of Quercetin on Injury to Indomethacin-Treated Human Embryonic Kidney 293 Cells. Life, 11(11), 1134.
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3

Apoptosis Pathway Evaluation Protocol

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AITC, 1,2-bis(2-aminophenoxy)ethane-N,N,N,N′-tetraacetic acid tetrakis(acetoxymethyl ester) (BAPTA/AM), calpeptin, 4′,6-diamidino-2-phenylindole (DAPI), N-acetylcysteine (NAC), propidium iodide (PI), and trypan blue were obtained from Sigma-Aldrich (Merck KGaA). 2′,7′-Dichlorodihydrofluorescein diacetate (H2DCFDA), 3,3′-dihexyloxacarbocyanine iodide [DiOC6(3 (link))], fetal bovine serum (FBS), 4-(6-acetoxymethoxy-2,7-dichloro-3-oxo-9-xanthenyl)-4′-methyl-2,2′(ethylenedioxy)dianiline-N, N,N,N′-tetraacetic acid tetrakis(acetoxymethyl) ester (fluo-3/AM), L-glutamine, Roswell Park Memorial Institute (RPMI)-1640 medium, penicillin, streptomycin, and trypsin-EDTA were obtained from Thermo Fisher Scientific, Inc. Caspase-3 and Caspase-9 Colorimetric Assay Kits were purchased from R&D Systems. Caspase-3 inhibitor Z-DEVD-FMK and caspase-9 inhibitor Z-LEHD-FMK were purchased from Merck Millipore. All primary antibodies were purchased from Cell Signaling Technology, Inc. or Santa Cruz Biotechnology, Inc. The goat anti-rabbit or anti-mouse immunoglobulin G (IgG) secondary antibodies conjugated with horseradish peroxidase (HRP) were also obtained from Cell Signaling Technology, Inc. Mitochondria/Cytosol Fractionation Kit was purchased from BioVision, Inc. The goat anti-mouse IgG-phycoerythrin (PE) secondary antibody was also obtained from Santa Cruz Biotechnology, Inc.
Chiang J.H., Tsai F.J., Hsu Y.M., Yin M.C., Chiu H.Y, & Yang J.S. (2020). Sensitivity of allyl isothiocyanate to induce apoptosis via ER stress and the mitochondrial pathway upon ROS production in colorectal adenocarcinoma cells. Oncology Reports, 44(4), 1415-1424.
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4

Caspase Activation in CAR Cells

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CAR cells were seeded at a density of 5×106 cells per 75T flask and incubated with 20 µg/ml cetuximab, 40 µM curcumin, or 20 µg/ml cetuximab and 40 µM curcumin for 24 h. The cell lysate was collected, and the cell fraction was analyzed for caspase-3/-9 activity using Caspase-3 and Caspase-9 Colorimetric Assay kits (R&D Systems, Inc., Minneapolis, MN, USA), according to the manufacturer's protocol.
Chen C.F., Lu C.C., Chiang J.H., Chiu H.Y., Yang J.S., Lee C.Y., Way T.D, & Huang H.J. (2018). Synergistic inhibitory effects of cetuximab and curcumin on human cisplatin-resistant oral cancer CAR cells through intrinsic apoptotic process. Oncology Letters, 16(5), 6323-6330.
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5

AITC Induces Caspase Activation in HT-29 Cells

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HT-29 cells (1×106 cells) in 75T flasks were exposed to 0, 5, 10, 15 and 20 µM of AITC for 24 h to assess the activities of caspase-9 and caspase-3, which were determined using Caspase-3 and Caspase-9 Colorimetric Assay Kits in accordance with the manufacturer's protocols (R&D Systems).
Chiang J.H., Tsai F.J., Hsu Y.M., Yin M.C., Chiu H.Y, & Yang J.S. (2020). Sensitivity of allyl isothiocyanate to induce apoptosis via ER stress and the mitochondrial pathway upon ROS production in colorectal adenocarcinoma cells. Oncology Reports, 44(4), 1415-1424.
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6

Curcumin-Induced Apoptosis in Cancer Cells

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Erbitux (the active ingredient of cetuximab) was provided by Hualien Tzu Chi Hospital (Taiwan) and originally purchased from Merck KGaA (Darmstadt, Germany). Dulbecco's modified Eagle's medium (DMEM), fetal bovine serum (FBS), L-glutamine and penicillin-streptomycin solution were purchased from HyClone (GE Healthcare, Logan, UT, USA). Caspase-3 and Caspase-9 colorimetric assay kits were sourced from R&D Systems (Minneapolis, MN, USA). All primary antibodies and anti-mouse/-rabbit immunoglobulin G (IgG) horseradish peroxidase (HRP)-conjugated antibodies were purchased from GeneTex (Hsinchu, Taiwan). Curcumin, Thiazolyl Blue Tetrazolium Bromide (MTT) and other reagents were of analytical grade from Sigma-Aldrich (Merck KGaA, Darmstadt Germany), unless otherwise stated.
Chen C.F., Lu C.C., Chiang J.H., Chiu H.Y., Yang J.S., Lee C.Y., Way T.D, & Huang H.J. (2018). Synergistic inhibitory effects of cetuximab and curcumin on human cisplatin-resistant oral cancer CAR cells through intrinsic apoptotic process. Oncology Letters, 16(5), 6323-6330.
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7

Caspase Activity Assay in PC-3 Cells

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After treatment, PC-3 cells were lyzed and activities of caspases were determined using caspase-3 and caspase-9 colorimetric assay kits in accordance with the manufacturer’s protocols (R&D Systems).
Xu H., Mao M., Zhao R, & Zhao Q. (2021). Enoxacin Exerts Anti-Tumor Effects Against Prostate Cancer Through Inducing Apoptosis. Technology in Cancer Research & Treatment, 20, 1533033821995284.
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8

Assessing Caspase-3 and -9 Activity in APG-Treated Cells

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AGS and SNU-638 cells (1 × 104 cells/well) were seeded into a 96-well plate with growth medium. To determine the caspase-3 and -9 activity (Minneapolis, MN, USA, R&D Systems Inc.) after APG treatment, cell lysates (50 μg proteins) were incubated to check relative caspase activity using Caspase-3 and Caspase-9 Colorimetric Assay Kits (Minneapolis, R&D Systems Inc.) following the manufacturer’s instructions.
Kim T.W, & Lee H.G. (2021). Apigenin Induces Autophagy and Cell Death by Targeting EZH2 under Hypoxia Conditions in Gastric Cancer Cells. International Journal of Molecular Sciences, 22(24), 13455.
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9

Apoptosis Induction and Oxidative Stress Assessment

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Culture media, growth supplements, penicillin, streptomycin and 0.25 % trypsin-EDTA solution were purchased from Gibco. Co (Germany). Annexin V-FITC apoptosis detection kit, propidium iodide (PI), MTT, JC-1 and dimethyl sulfoxide (DMSO) were from Sigma-Aldrich (Munich, Germany). Caspase-3 and caspase-9 colorimetric assay kits were obtained from R&D systems Co. (Minneapolis, United States). Caspase-6 colorimetric assay kit was from BioVision (BioVision, Inc. Milpitas, CA USA). Mouse monoclonal anti-p53 antibody, anti-Bcl-2 antibody, anti-Bax, antibody, and anti-mouse IgG-HRP antibody were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Fluorescent Reactive Oxygen Species detection kit was obtained from Marker Gene Technologies (MGT, Inc., USA).
Fallahian F., Aghaei M., Abdolmohammadi M., & Hamzeloo-Moghadam M. (2016). Molecular mechanism of apoptosis induction by Gaillardin, a sesquiterpene lactone, in breast cancer cell lines. Cell Biology and Toxicology, 31(6), 295-305.
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