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Apo one caspase 3 7 reagent

Manufactured by Promega
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The Apo-ONE Caspase 3/7 reagent is a fluorescent-based assay kit used to measure the activity of caspase-3 and caspase-7, two key enzymes involved in the process of apoptosis, or programmed cell death. The reagent includes a profluorescent substrate that, when cleaved by the target caspases, releases a fluorescent product that can be quantified using a fluorometer or plate reader.

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Lab products found in correlation

Polarstar optima microplate reader, by BMG Labtech (1 mentions) Caspase 3 7 activity, by Promega (1 mentions)

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Apo-ONE (13 citations) Caspase-3 (7 citations)

4 protocols using apo one caspase 3 7 reagent

1

Caspase 3/7 Apoptosis Assay

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Cells were seeded in a 96-well plate at a concentration of 8 × 103 to 10 × 103 cells per well 40 h prior to treatment. After treatment, 50 µL of Apo-ONE Caspase 3/7 reagent (Promega, no. G7790) was added to 70 µL of medium in each well. Plates were rocked in the dark for 20 min and then frozen at −80°C before being thawed to achieve complete cell lysis. Plates were analyzed on a POLARstar Optima microplate reader (BMG Labtech) at 485-nm excitation and 520 emission.
O'Neill K.L., Huang K., Zhang J., Chen Y, & Luo X. (2016). Inactivation of prosurvival Bcl-2 proteins activates Bax/Bak through the outer mitochondrial membrane. Genes & Development, 30(8), 973-988.
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2

Apoptosis induction in prostate cancer cells

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LNCaP, C4-2B, ARCaP, VCaP, PC3 and DU145 were cultured in white-walled 96-well plates at a concentration of 1×104 per 50μl per well. Cells were treated with 50μl DI176E (0.1μg/ml, 1μg/ml, 10μg/ml and 100μg/ml final concentrations) or vehicle or etoposide (100nM final concentration) (Sigma) as a positive control and were allowed to grow for 24 hours at which point 100ul of Apo-ONE Caspase-3/7 Reagent (Promega, Madison, WI) was added to each well and the cells were incubated for extended periods (>4 hours). Absorbance was determined at 499 and 521 nm using a plate reader (Multi-Mode Microplate Reader, SpectraMax M5, Molecular Devices MDS Analytical Technologies).
Jiang Y., Dai J., Yao Z., Shelley G, & Keller E.T. (2017). Abituzumab Targeting of αV-class Integrins Inhibits Prostate Cancer Progression. Molecular cancer research : MCR, 15(7), 875-883.
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3

Apoptosis Induction Assay with PAC-320

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LNCaP or DU145 cells were plated at 2 × 103 cells per well on 96-well plates. At 24 h after treatment with PAC-320, the Apo-ONE caspase 3/7 Reagent (Promega, Madison/Wisconsin) was added. Cells were incubated for 1 h at room temperature prior to record of the fluorescence (485Ex/527Em).
Dong Z., Yang Y., Liu S., Lu J., Huang B, & Zhang Y. (2017). HDAC inhibitor PAC-320 induces G2/M cell cycle arrest and apoptosis in human prostate cancer. Oncotarget, 9(1), 512-523.
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4

Caspase 3/7 Activity Apoptosis Assay

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The apoptosis assay based on the measurement of Caspase 3/7 activity was performed according to the manufacturer’s instructions (Promega). Briefly, 100 μL of Apo-ONE Caspase 3/7 reagent (Promega) was added into each well of white-walled 96-well (Nunc) plate and incubated on a shaker at 300–500 rpm for 5 min followed by one hour incubation at room temperature. The luminescence of each sample was measured using a luminometer.
Murtaza M., Shan J., Matigian N., Todorovic M., Cook A.L., Ravishankar S., Dong L.F., Neuzil J., Silburn P., Mackay-Sim A., Mellick G.D, & Wood S.A. (2016). Rotenone Susceptibility Phenotype in Olfactory Derived Patient Cells as a Model of Idiopathic Parkinson’s Disease. PLoS ONE, 11(4), e0154544.
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