Mycoplasma tested human umbilical vein endothelial cells (HUVECs; Commercially obtained from Lonza #CC‐2935) were cultured in EBM‐2 Basal Media (#00190860, #cc‐4176, Lonza, mycoplasma free) with EGM‐2 Single Quots supplements (Lonza #cc‐4176) on 0.2% gelatin (Sigma Aldrich #G1393)‐coated 96‐well plates (5,000 cells/well) or 6‐well plates (150,000 cells/well). After obtaining stable culture conditions, HUVECs were incubated with culture supernatants collected from three groups of mouse myofibroblasts: (i) untreated myofibroblasts (TurboFect), (ii) control siRNA‐treated myofibroblasts, and (iii) myofibroblasts treated with siRNA against PDGF‐Rα. For caspase activity assay, incubation with 40 μg/ml anti‐VEGF (C‐1) antibody (#sc‐7269, Santa Cruz) served as a positive control. Caspase activity was assessed after 6 h using the Caspase‐Glo 3/7 assay kit (#G8091, Promega). After 6 h of incubation, caspase activity was assessed using the Caspase‐Glo 3/7 assay kit (Promega GmbH, Germany #G8091) according to the manufacturer's instructions. Cells plated in 6‐well plate were lysed after 6‐h incubation, and lysate was processed for immunoblot analysis with cleaved caspase‐9 (Cell Signaling Technologies #9509) and eNOS (Cell Signaling Technologies #880) protein.
Caspase glo 3 7 assay kit
Manufactured by Promega
Sourced in United States, United Kingdom, Germany, France
The Caspase-Glo 3/7 assay kit is a luminescent-based assay that measures the activity of caspase-3 and caspase-7 enzymes. The kit uses a proluminescent caspase-3/7 substrate, which can be cleaved by these enzymes, resulting in the release of a signal that is proportional to the amount of caspase activity present in the sample.
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Lab products found in correlation
Glomax multi detection system, by Promega (12 mentions)
Celltiter glo luminescent cell viability assay kit, by Promega (10 mentions)
Caspase glo 9 assay kit, by Promega (8 mentions)
Luminoskan ascent, by Thermo Fisher Scientific (6 mentions)
Epoch microplate reader, by Agilent Technologies (5 mentions)
Fluostar optima, by BMG Labtech (5 mentions)
Synergy 2 multi mode microplate reader, by Agilent Technologies (5 mentions)
Glomax luminometer, by Promega (5 mentions)
Glomax 20 20 luminometer, by Promega (5 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (5 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Fluostar omega, by BMG Labtech (4 mentions)
Synergy ht, by Agilent Technologies (4 mentions)
Microplate reader, by Agilent Technologies (4 mentions)
Fluostar omega microplate reader, by BMG Labtech (4 mentions)
Multimode mithras lb 940, by Berthold Technologies (4 mentions)
Synergy ht multi mode microplate reader, by Agilent Technologies (4 mentions)
Infinite 200 pro, by Tecan (4 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (4 mentions)
575 protocols using caspase glo 3 7 assay kit
1
HUVEC Caspase Activity Assay
2
Caspase 3/7 Activity in CLL Cells
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CLL cell Caspase 3/7 activity after TRIP13-KD and NC group were performed following the manual of Promega Caspase-Glo® 3/7 assay kit (Promega, USA). Briefly, TRIP13-KD and NC lentiverus infected cells were planted in 96 well plates for 3 days and 1 × 104 cells were isolated and added to 100 μl Caspase-Glo buffer. After 300-500 rpm for 30 min, the samples were incubated in room temperature for 0.5 hours and assayed by a microplate reader.
3
Caspase-3/7 Activation in Melanoma
4
Caspase 3/7 Activity Assay in CLL Cells
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Caspase 3/7 activity in CLL cells was detected using the Promega Caspase-Glo 3/7 assay kit (Promega). Briefly, cells infected with shctrl or shKIAA0101 lentiviruses were plated into 96-well plates and incubated for 5 days. Next, cells were collected and cell suspensions (1×104 cells/well) were reacted with 100 μL of Caspase-Glo reaction solution. After shaking for 30 min at 300–500 rpm, the reaction system was incubated for 2 h at room temperature. Finally, signal intensity was determined using a microplate reader.
5
Detecting Adipocyte Apoptosis by Multiple Assays
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Apoptosis in adipocytes was observed and detected by various methods, including caspase 3/7 activation analysis, TUNEL assay, and flow cytometry. Caspase 3/7 activity was quantitated by using a Caspase-Glo® 3/7 Assay Kit (Promega, CA, USA) according to the manufacturer's instructions. A DeadEnd™ Fluorometric TUNEL System (Promega) was used to observe and measure the adipocyte apoptosis. The number of apoptotic adipocytes was quantitated by flow cytometry assay (Gallios, Beckman Coulter, USA) with the Annexin V/PI double-staining method according to the manufacturer's instructions. (The Annexin V/PI apoptosis assay kit was purchased from Life Technologies, USA).
6
Cell Proliferation and Apoptosis Assay
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CellTiter 96® Aqueous One Solution Cell Proliferation assay (MTS) and Caspase-Glo 3/7 Assay Kit were both obtained from Promega Corporation (Madison, WI, USA). Hoechst 33342 was purchased from Sigma-Aldrich (St. Louis, MO, USA). Dead Cell Apoptosis Kit with Annexin V Alexa Fluor® 488 and propidium iodide (PI), and fetal bovine serum (FBS) were from Invitrogen (Grand Island, NY, USA). Quantikine Human VEGF Immunoassay Kit was purchased from R&D Systems (Minneapolis, MN, USA). Primary antibodies for Bad (C-7) (#8044), Bcl-xL (H-5) (#8392), and GAPDH (0411) (#47724) were from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Bcl-2 (#2872S), HIF-1α (#14179S), hVEGF (#8065), FasL (#4273S), FADD (#2782S), DR5 (#3696S), monoclonal pro-caspase-9 (C9) (#9508S), pro-caspase-3 (#9662S), and pro-caspase-7 (#9492S) antibodies and horseradish peroxidase-conjugated secondary antibody were purchased from Cell Signaling Technology, Inc. (Danvers, MA, USA). Antibody HIF-1β (#611079) was from BD Biosciences (San Jose, CA, USA). The BCA Protein Assay Kit was obtained from Thermo Fisher Scientific (Waltham, MA, USA). HaltTM Protease and Phosphatase Inhibitor Single-Use Cocktail were purchased from Life Technologies (Grand Island, NY, USA). EDTA solution was form Thermo Scientific (Rockford, IL, USA).
7
Caspase-3/7 Activity Assay in Cells
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The enzymatic activity of caspase-3/7 was determined using the Caspase-Glo 3/7 assay kit following the manufacturer’s instructions (Promega, Madison, WI, United States), as previously reported (Konno et al., 2014 (link)). Briefly, 3 × 103 cells were plated in triplicates in 96-well plates and transfected as indicated. An equal amount of Caspase-Glo 3/7 substrate was added to the culture and subsequently incubated for 3 h. Following incubation, caspase-3/7 activities were evaluated as an indicator of cell apoptosis using a GloMax-96 Microplate luminometer (Promega). The results were shown as the fold change relative to the control cells.
8
Caspase-3/7 Activity Assay Protocol
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To determine the induction of apoptosis, the activity of caspase-3/7 was measured using the Caspase-Glo 3/7 assay kit (Promega), in accordance with the manufacturer’s instructions.
9
Quantifying Cell Apoptosis via Caspase Assay
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To assess cell apoptotic rate, we utilized Caspase-Glo® 3/7 Assay kit (Promega) for their value test. All steps were performed according to manufacturer’s instructions. The activities of caspase-3/7 were recorded following luminescence reading.
10
Apoptosis Measurement in DLBCL Cells
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Apoptosis was assessed using the luminescence-based Caspase-Glo 3/7 assay kit (Promega) according to manufacturers' instructions. Exponentially growing DLBCL cell lines were seeded into 96-well plates and incubated in the presence of previously determined saturating concentrations of IMGN529 or rituximab, both alone and in combination, for 24 hours. Caspase-Glo 3/7 reagent was added (50% v/v) to the cells, and plates were incubated for 30 minutes before luminescence detection in a Victor3 microplate reader (PerkinElmer). Additional caspase3/7 activation studies were performed combining IMGN529 with ofatumumab, obinutuzumab, K7153A, and IgG1-SMCC-DM1 in the U-2932 and SU-DHL-4 cell lines using the same protocol.
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