Structured Review

Promega caspase activity
GBM cells display augmented apoptotic response upon chaetocin and TRAIL combinatorial treatment. a <t>Caspase-3/7</t> activity analyses of U87MG cells upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment. Data were normalized to untreated control cells. b Western blot analyses of U87MG cells for cleaved Casp8, Bid, t-Bid, Casp3 and PARP after pretreatment with chaetocin (100 nM for 24 h) followed by 6 h TRAIL (100 ng/ml) treatment. α - tubulin was shown as protein loading control. c Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay on U87MG cells showing increased DNA fragmentation upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment for 24 h. Blue: DAPI staining nuclei, Green: TUNEL (+) cells. Scale bar: 100 µm. d Quantification of TUNEL staining by ImageJ program through counting TUNEL (+) cells with green fluorescence. e YO-PRO-1 and PI staining upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment of U87MG cells for 24 h. Green: YO-PRO-1 staining apoptotic cells, Red: PI staining dead/necrotic cells. Scale bar: 200 µm. f Quantification of YO-PRO-1/PI staining by ImageJ program through counting green and red fluorescence positive cells. g Flow cytometric analysis of Annexin V/PI stained U87MG cells upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment for 24 h. h Quantification of flow cytometry data showing marked increase in apoptotic cell populations upon combinatorial treatment. Data were normalized to total number of cells under each condition. i Cell viability analysis U87MG cells pretreated with caspase inhibitors (20 µM for 24 h) followed by chaetocin (100 nM) and TRAIL (100 ng/ml) treatment for 24 h in presence of inhibitors. Z-FA-FMK: negative control, Z-VAD-FMK: general caspase inhibitor. j Western blot analyses of U87MG cells showing individual stable CRISPR knockouts of DR5, BID, and Casp8, as well as double knock out Casp3/7 genes. GFP targeting guide RNA (g-GFP) was used as negative control for CRISPR assay. α-tubulin was shown as protein loading control. k Viability analysis of CRISPR edited U87MG cells upon combinatorial treatment with chaetocin (100 nM) and TRAIL (100 ng/ml) for 24 h. Data were normalized to untreated control. ((*) and (***) denote P
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Images

1) Product Images from "The fungal metabolite chaetocin is a sensitizer for pro-apoptotic therapies in glioblastoma"

Article Title: The fungal metabolite chaetocin is a sensitizer for pro-apoptotic therapies in glioblastoma

Journal: Cell Death & Disease

doi: 10.1038/s41419-019-2107-y

GBM cells display augmented apoptotic response upon chaetocin and TRAIL combinatorial treatment. a Caspase-3/7 activity analyses of U87MG cells upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment. Data were normalized to untreated control cells. b Western blot analyses of U87MG cells for cleaved Casp8, Bid, t-Bid, Casp3 and PARP after pretreatment with chaetocin (100 nM for 24 h) followed by 6 h TRAIL (100 ng/ml) treatment. α - tubulin was shown as protein loading control. c Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay on U87MG cells showing increased DNA fragmentation upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment for 24 h. Blue: DAPI staining nuclei, Green: TUNEL (+) cells. Scale bar: 100 µm. d Quantification of TUNEL staining by ImageJ program through counting TUNEL (+) cells with green fluorescence. e YO-PRO-1 and PI staining upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment of U87MG cells for 24 h. Green: YO-PRO-1 staining apoptotic cells, Red: PI staining dead/necrotic cells. Scale bar: 200 µm. f Quantification of YO-PRO-1/PI staining by ImageJ program through counting green and red fluorescence positive cells. g Flow cytometric analysis of Annexin V/PI stained U87MG cells upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment for 24 h. h Quantification of flow cytometry data showing marked increase in apoptotic cell populations upon combinatorial treatment. Data were normalized to total number of cells under each condition. i Cell viability analysis U87MG cells pretreated with caspase inhibitors (20 µM for 24 h) followed by chaetocin (100 nM) and TRAIL (100 ng/ml) treatment for 24 h in presence of inhibitors. Z-FA-FMK: negative control, Z-VAD-FMK: general caspase inhibitor. j Western blot analyses of U87MG cells showing individual stable CRISPR knockouts of DR5, BID, and Casp8, as well as double knock out Casp3/7 genes. GFP targeting guide RNA (g-GFP) was used as negative control for CRISPR assay. α-tubulin was shown as protein loading control. k Viability analysis of CRISPR edited U87MG cells upon combinatorial treatment with chaetocin (100 nM) and TRAIL (100 ng/ml) for 24 h. Data were normalized to untreated control. ((*) and (***) denote P
Figure Legend Snippet: GBM cells display augmented apoptotic response upon chaetocin and TRAIL combinatorial treatment. a Caspase-3/7 activity analyses of U87MG cells upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment. Data were normalized to untreated control cells. b Western blot analyses of U87MG cells for cleaved Casp8, Bid, t-Bid, Casp3 and PARP after pretreatment with chaetocin (100 nM for 24 h) followed by 6 h TRAIL (100 ng/ml) treatment. α - tubulin was shown as protein loading control. c Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay on U87MG cells showing increased DNA fragmentation upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment for 24 h. Blue: DAPI staining nuclei, Green: TUNEL (+) cells. Scale bar: 100 µm. d Quantification of TUNEL staining by ImageJ program through counting TUNEL (+) cells with green fluorescence. e YO-PRO-1 and PI staining upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment of U87MG cells for 24 h. Green: YO-PRO-1 staining apoptotic cells, Red: PI staining dead/necrotic cells. Scale bar: 200 µm. f Quantification of YO-PRO-1/PI staining by ImageJ program through counting green and red fluorescence positive cells. g Flow cytometric analysis of Annexin V/PI stained U87MG cells upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment for 24 h. h Quantification of flow cytometry data showing marked increase in apoptotic cell populations upon combinatorial treatment. Data were normalized to total number of cells under each condition. i Cell viability analysis U87MG cells pretreated with caspase inhibitors (20 µM for 24 h) followed by chaetocin (100 nM) and TRAIL (100 ng/ml) treatment for 24 h in presence of inhibitors. Z-FA-FMK: negative control, Z-VAD-FMK: general caspase inhibitor. j Western blot analyses of U87MG cells showing individual stable CRISPR knockouts of DR5, BID, and Casp8, as well as double knock out Casp3/7 genes. GFP targeting guide RNA (g-GFP) was used as negative control for CRISPR assay. α-tubulin was shown as protein loading control. k Viability analysis of CRISPR edited U87MG cells upon combinatorial treatment with chaetocin (100 nM) and TRAIL (100 ng/ml) for 24 h. Data were normalized to untreated control. ((*) and (***) denote P

Techniques Used: Activity Assay, Western Blot, TUNEL Assay, Staining, Fluorescence, Flow Cytometry, Cytometry, Negative Control, CRISPR, Knock-Out

2) Product Images from "The fungal metabolite chaetocin is a sensitizer for pro-apoptotic therapies in glioblastoma"

Article Title: The fungal metabolite chaetocin is a sensitizer for pro-apoptotic therapies in glioblastoma

Journal: Cell Death & Disease

doi: 10.1038/s41419-019-2107-y

GBM cells display augmented apoptotic response upon chaetocin and TRAIL combinatorial treatment. a Caspase-3/7 activity analyses of U87MG cells upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment. Data were normalized to untreated control cells. b Western blot analyses of U87MG cells for cleaved Casp8, Bid, t-Bid, Casp3 and PARP after pretreatment with chaetocin (100 nM for 24 h) followed by 6 h TRAIL (100 ng/ml) treatment. α - tubulin was shown as protein loading control. c Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay on U87MG cells showing increased DNA fragmentation upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment for 24 h. Blue: DAPI staining nuclei, Green: TUNEL (+) cells. Scale bar: 100 µm. d Quantification of TUNEL staining by ImageJ program through counting TUNEL (+) cells with green fluorescence. e YO-PRO-1 and PI staining upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment of U87MG cells for 24 h. Green: YO-PRO-1 staining apoptotic cells, Red: PI staining dead/necrotic cells. Scale bar: 200 µm. f Quantification of YO-PRO-1/PI staining by ImageJ program through counting green and red fluorescence positive cells. g Flow cytometric analysis of Annexin V/PI stained U87MG cells upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment for 24 h. h Quantification of flow cytometry data showing marked increase in apoptotic cell populations upon combinatorial treatment. Data were normalized to total number of cells under each condition. i Cell viability analysis U87MG cells pretreated with caspase inhibitors (20 µM for 24 h) followed by chaetocin (100 nM) and TRAIL (100 ng/ml) treatment for 24 h in presence of inhibitors. Z-FA-FMK: negative control, Z-VAD-FMK: general caspase inhibitor. j Western blot analyses of U87MG cells showing individual stable CRISPR knockouts of DR5, BID, and Casp8, as well as double knock out Casp3/7 genes. GFP targeting guide RNA (g-GFP) was used as negative control for CRISPR assay. α-tubulin was shown as protein loading control. k Viability analysis of CRISPR edited U87MG cells upon combinatorial treatment with chaetocin (100 nM) and TRAIL (100 ng/ml) for 24 h. Data were normalized to untreated control. ((*) and (***) denote P
Figure Legend Snippet: GBM cells display augmented apoptotic response upon chaetocin and TRAIL combinatorial treatment. a Caspase-3/7 activity analyses of U87MG cells upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment. Data were normalized to untreated control cells. b Western blot analyses of U87MG cells for cleaved Casp8, Bid, t-Bid, Casp3 and PARP after pretreatment with chaetocin (100 nM for 24 h) followed by 6 h TRAIL (100 ng/ml) treatment. α - tubulin was shown as protein loading control. c Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay on U87MG cells showing increased DNA fragmentation upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment for 24 h. Blue: DAPI staining nuclei, Green: TUNEL (+) cells. Scale bar: 100 µm. d Quantification of TUNEL staining by ImageJ program through counting TUNEL (+) cells with green fluorescence. e YO-PRO-1 and PI staining upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment of U87MG cells for 24 h. Green: YO-PRO-1 staining apoptotic cells, Red: PI staining dead/necrotic cells. Scale bar: 200 µm. f Quantification of YO-PRO-1/PI staining by ImageJ program through counting green and red fluorescence positive cells. g Flow cytometric analysis of Annexin V/PI stained U87MG cells upon chaetocin (100 nM) and TRAIL (100 ng/ml) combinatorial treatment for 24 h. h Quantification of flow cytometry data showing marked increase in apoptotic cell populations upon combinatorial treatment. Data were normalized to total number of cells under each condition. i Cell viability analysis U87MG cells pretreated with caspase inhibitors (20 µM for 24 h) followed by chaetocin (100 nM) and TRAIL (100 ng/ml) treatment for 24 h in presence of inhibitors. Z-FA-FMK: negative control, Z-VAD-FMK: general caspase inhibitor. j Western blot analyses of U87MG cells showing individual stable CRISPR knockouts of DR5, BID, and Casp8, as well as double knock out Casp3/7 genes. GFP targeting guide RNA (g-GFP) was used as negative control for CRISPR assay. α-tubulin was shown as protein loading control. k Viability analysis of CRISPR edited U87MG cells upon combinatorial treatment with chaetocin (100 nM) and TRAIL (100 ng/ml) for 24 h. Data were normalized to untreated control. ((*) and (***) denote P

Techniques Used: Activity Assay, Western Blot, TUNEL Assay, Staining, Fluorescence, Flow Cytometry, Cytometry, Negative Control, CRISPR, Knock-Out

3) Product Images from "MTT ASSAYS CANNOT BE UTILIZED TO STUDY THE EFFECTS OF STI571/GLEEVEC ON THE VIABILITY OF SOLID TUMOR CELL LINES"

Article Title: MTT ASSAYS CANNOT BE UTILIZED TO STUDY THE EFFECTS OF STI571/GLEEVEC ON THE VIABILITY OF SOLID TUMOR CELL LINES

Journal: Cancer chemotherapy and pharmacology

doi: 10.1007/s00280-009-1004-y

STI571 interferes with the ability of the MTT assay to measure cell viability
Figure Legend Snippet: STI571 interferes with the ability of the MTT assay to measure cell viability

Techniques Used: MTT Assay

STI571 inhibits cell growth, proliferation, and induces apoptosis of cells containing highly active Abl kinases
Figure Legend Snippet: STI571 inhibits cell growth, proliferation, and induces apoptosis of cells containing highly active Abl kinases

Techniques Used:

4) Product Images from "Identification of SERPINE1 as a Regulator of Glioblastoma Cell Dispersal with Transcriptome Profiling"

Article Title: Identification of SERPINE1 as a Regulator of Glioblastoma Cell Dispersal with Transcriptome Profiling

Journal: Cancers

doi: 10.3390/cancers11111651

SERPINE1 knock-down reduces GBM dispersal ( A ) qRT-PCR analysis of SERPINE1 expression levels after shRNA knock-down; ( B ) SERPINE1 protein levels after shRNA knock-down; ( C ) dispersal assay that shows SERPINE1 knock-down reduces dispersal of U373 and A172 spheroids significantly ( n = 24 spheroids for each condition, scale bar: 200 µm); ( D ) dispersal assays that shows chemical inhibitor of SERPINE1, tiplaxtinin, reduces dispersal of U373 spheroids ( n = 12 spheroids for each condition, scale bar: 200 µm); ( E ) wound healing analysis of the effect of SERPINE1 knock-down ( n = 35 areas for each condition, scale bar: 200 µm); ( F ) cell viability analysis of the effects of SERPINE1 knock-down in U373 and A172 cell growth. (*, ** and *** denote p
Figure Legend Snippet: SERPINE1 knock-down reduces GBM dispersal ( A ) qRT-PCR analysis of SERPINE1 expression levels after shRNA knock-down; ( B ) SERPINE1 protein levels after shRNA knock-down; ( C ) dispersal assay that shows SERPINE1 knock-down reduces dispersal of U373 and A172 spheroids significantly ( n = 24 spheroids for each condition, scale bar: 200 µm); ( D ) dispersal assays that shows chemical inhibitor of SERPINE1, tiplaxtinin, reduces dispersal of U373 spheroids ( n = 12 spheroids for each condition, scale bar: 200 µm); ( E ) wound healing analysis of the effect of SERPINE1 knock-down ( n = 35 areas for each condition, scale bar: 200 µm); ( F ) cell viability analysis of the effects of SERPINE1 knock-down in U373 and A172 cell growth. (*, ** and *** denote p

Techniques Used: Quantitative RT-PCR, Expressing, shRNA

SERPINE1 expression is correlated with poor prognosis and its silencing in a clinically-relevant model reduces cell viability and dispersal. ( A ) TCGA survival data plotted for high/low SERPINE1 expressing glioma patients. ( B ) SERPINE1 expression correlation with LGG samples and GBM samples; ( C ) SERPINE1 expression correlation with GBM subtypes; ( D ) SERPINE 1 mRNA and protein levels after shRNA knock-down in GBM8 cells; ( E ) cell viability assays that show SERPINE1 knockdown slows down GBM8 cell proliferation; ( F ) live cell imaging and analysis of shControl and shSERPINE1 spheroids. SERPINE1 knockdown reduces dispersal of GBM8 spheroids (video for five hours of dispersal, images taken every 5 min, scale bar: 200 µm, n = 10 spheroids for each condition); ( G ) live cell imaging analysis of DMSO- or Tiplaxtinin-treated GBM8 spheroids. (video for five hours of dispersal, images taken in every 60 min, magnification is 10×, n = 12 spheroids for each condition); ( H ) SERPINE1 knock-down reduces the number of attached cells in different time points (three wells analyzed for each condition and each time point). (*, ** and *** denote p
Figure Legend Snippet: SERPINE1 expression is correlated with poor prognosis and its silencing in a clinically-relevant model reduces cell viability and dispersal. ( A ) TCGA survival data plotted for high/low SERPINE1 expressing glioma patients. ( B ) SERPINE1 expression correlation with LGG samples and GBM samples; ( C ) SERPINE1 expression correlation with GBM subtypes; ( D ) SERPINE 1 mRNA and protein levels after shRNA knock-down in GBM8 cells; ( E ) cell viability assays that show SERPINE1 knockdown slows down GBM8 cell proliferation; ( F ) live cell imaging and analysis of shControl and shSERPINE1 spheroids. SERPINE1 knockdown reduces dispersal of GBM8 spheroids (video for five hours of dispersal, images taken every 5 min, scale bar: 200 µm, n = 10 spheroids for each condition); ( G ) live cell imaging analysis of DMSO- or Tiplaxtinin-treated GBM8 spheroids. (video for five hours of dispersal, images taken in every 60 min, magnification is 10×, n = 12 spheroids for each condition); ( H ) SERPINE1 knock-down reduces the number of attached cells in different time points (three wells analyzed for each condition and each time point). (*, ** and *** denote p

Techniques Used: Expressing, shRNA, Live Cell Imaging

5) Product Images from "MTT ASSAYS CANNOT BE UTILIZED TO STUDY THE EFFECTS OF STI571/GLEEVEC ON THE VIABILITY OF SOLID TUMOR CELL LINES"

Article Title: MTT ASSAYS CANNOT BE UTILIZED TO STUDY THE EFFECTS OF STI571/GLEEVEC ON THE VIABILITY OF SOLID TUMOR CELL LINES

Journal: Cancer chemotherapy and pharmacology

doi: 10.1007/s00280-009-1004-y

STI571 interferes with the ability of the MTT assay to measure cell viability
Figure Legend Snippet: STI571 interferes with the ability of the MTT assay to measure cell viability

Techniques Used: MTT Assay

STI571 inhibits cell growth, proliferation, and induces apoptosis of cells containing highly active Abl kinases
Figure Legend Snippet: STI571 inhibits cell growth, proliferation, and induces apoptosis of cells containing highly active Abl kinases

Techniques Used:

Related Articles

Irradiation:

Article Title: Near‐infrared photoimmunotherapy targeting EGFR—Shedding new light on glioblastoma treatment
Article Snippet: Wells were irradiated in groups of nine using a red LED L690‐66‐60 (Marubeni, Tokyo, Japan). .. Cell response to PIT was evaluated by the CellTiter‐Glo® luminescent cell viability assay (Promega, Madison, WI) 24 or 96 h post‐irradiation in the 2D and 3D cell cultures.

Negative Control:

Article Title: WNK1 kinase and its partners Akt, SGK1 and NBC-family Na+/HCO3− cotransporters are potential therapeutic targets for glioblastoma stem-like cells linked to Bisacodyl signaling
Article Snippet: Negative control wells contained cells treated with DMSO (1%; vehicle). .. For macro-spheres, cell viability assays were performed with CellTiter-Glo® 3D cell viability assay (Promega).

In Vitro:

Article Title: Near‐infrared photoimmunotherapy targeting EGFR—Shedding new light on glioblastoma treatment
Article Snippet: Paragraph title: Photoimmunotherapy in vitro studies ... Cell response to PIT was evaluated by the CellTiter‐Glo® luminescent cell viability assay (Promega, Madison, WI) 24 or 96 h post‐irradiation in the 2D and 3D cell cultures.

Article Title: Engineering human cell spheroids to model embryonic tissue fusion in vitro
Article Snippet: Paragraph title: Establishment of morphogenetic fusion model in vitro ... On fusion day 4, medium in each well was replaced with 100 μL per well of CellTiter Glo 3D reagent (Promega), and spheroids was incubated for 20 min on an orbital shaker at 37°C.

Transfection:

Article Title: Large-scale data integration framework provides a comprehensive view on glioblastoma multiforme
Article Snippet: .. Cell proliferation was assayed 72 h after transfection using CellTiter-Glo Cell Viability assay. ..

Article Title: Lipid-mRNA Nanoparticle Designed to Enhance Intracellular Delivery Mediated by Shock Waves
Article Snippet: Cell viability was measured using CellTiter-Glo cell viability assay (Promega G7573). .. Cells were cultured in 96-well plates and, after transfection, were incubated for 24 h, the medium was removed and 100 μL of the CellTiter-Glo reagent was added to each well and mixed by gentle shaking.

Serial Dilution:

Article Title: The Hsp70 co-chaperone Ydj1/HDJ2 regulates ribonucleotide reductase activity
Article Snippet: Similarly, cells were treated with either DMSO (control) or 40 μM of 116-9e in combination with a ten-fold serial dilution of either HU (400μM to 1.56 μM) or triapine (250μM to 0.0005 μM). .. After 72 h, cell viability was measured using Promega CellTiter-Glo cell viability assay on a Synergy H1 plate reader.

Cell Culture:

Article Title: The Hsp70 co-chaperone Ydj1/HDJ2 regulates ribonucleotide reductase activity
Article Snippet: Paragraph title: Mammalian cell culture and drug treatment ... After 72 h, cell viability was measured using Promega CellTiter-Glo cell viability assay on a Synergy H1 plate reader.

Article Title: Lipid-mRNA Nanoparticle Designed to Enhance Intracellular Delivery Mediated by Shock Waves
Article Snippet: Cell viability was measured using CellTiter-Glo cell viability assay (Promega G7573). .. Cells were cultured in 96-well plates and, after transfection, were incubated for 24 h, the medium was removed and 100 μL of the CellTiter-Glo reagent was added to each well and mixed by gentle shaking.

Article Title: Hedgehog signaling promotes sorafenib resistance in hepatocellular carcinoma patient-derived organoids
Article Snippet: Drug treatment HCC PDOs were seeded in 10 μl of GFR Matrigel matrix droplets in a 96-well plate (Corning) and cultured for 6 days. .. Cell viability was detected using CellTiter-Glo 3D reagent (Promega,G9681).

Article Title: A Novel 3-dimensional High Throughput Screening Approach Identifies Inducers of a Mutant KRAS Selective Lethal Phenotype
Article Snippet: Paragraph title: 3D Cell Culture and 3D Luminescent Proliferation Assay ... Plates were incubated for an additional 24 hours (for a total of 48 hours) under the same atmosphere and then treated with 20 μL per well of CellTiter-Glo 3D (Part#G9683, Promega Corp., Madison, WI).

Cytometry:

Article Title: Near‐infrared photoimmunotherapy targeting EGFR—Shedding new light on glioblastoma treatment
Article Snippet: Cell response to PIT was evaluated by the CellTiter‐Glo® luminescent cell viability assay (Promega, Madison, WI) 24 or 96 h post‐irradiation in the 2D and 3D cell cultures. .. In addition, formation and growth of 3D spheroids was monitored daily by the Celigo® image cytometry system (Nexcelom Bioscience, Lawrence, MA).

Microscopy:

Article Title: A Novel 3-dimensional High Throughput Screening Approach Identifies Inducers of a Mutant KRAS Selective Lethal Phenotype
Article Snippet: This allowed for spheroid formation, which was verified using a bright field microscope (Thermo Fisher). .. Plates were incubated for an additional 24 hours (for a total of 48 hours) under the same atmosphere and then treated with 20 μL per well of CellTiter-Glo 3D (Part#G9683, Promega Corp., Madison, WI).

Concentration Assay:

Article Title: Engineering human cell spheroids to model embryonic tissue fusion in vitro
Article Snippet: Alternatively, spheroids were treated at day 0 with various concentrations of recombinant human EGF (BioLegend) that was reconstituted at 10 μg/mL in sterile 1% BSA in DPBS, diluted with 1% BSA in DPBS, and added to spheroids at either 2, 10, or 50 ng/mL final concentration in CnT-PR-CC with 1% antibiotic-antimycotic, with the vehicle control containing the same dilution of 1% BSA in DPBS. .. On fusion day 4, medium in each well was replaced with 100 μL per well of CellTiter Glo 3D reagent (Promega), and spheroids was incubated for 20 min on an orbital shaker at 37°C.

Article Title: A Novel 3-dimensional High Throughput Screening Approach Identifies Inducers of a Mutant KRAS Selective Lethal Phenotype
Article Snippet: Cells were then suspended to the appropriate concentration for dispensing into Corning 384-well format 3D spheroid culture plates (part#3830, Corning Inc., NY). .. Plates were incubated for an additional 24 hours (for a total of 48 hours) under the same atmosphere and then treated with 20 μL per well of CellTiter-Glo 3D (Part#G9683, Promega Corp., Madison, WI).

Incubation:

Article Title: Engineering human cell spheroids to model embryonic tissue fusion in vitro
Article Snippet: .. On fusion day 4, medium in each well was replaced with 100 μL per well of CellTiter Glo 3D reagent (Promega), and spheroids was incubated for 20 min on an orbital shaker at 37°C. .. Reagent from each well was then transferred to a white opaque 96 well plate (Falcon) and read on a luminescence plate reader with 10 s integration time per well (Clariostar BMG Labtech).

Article Title: Lipid-mRNA Nanoparticle Designed to Enhance Intracellular Delivery Mediated by Shock Waves
Article Snippet: Cell viability was measured using CellTiter-Glo cell viability assay (Promega G7573). .. Cells were cultured in 96-well plates and, after transfection, were incubated for 24 h, the medium was removed and 100 μL of the CellTiter-Glo reagent was added to each well and mixed by gentle shaking.

Article Title: A Novel 3-dimensional High Throughput Screening Approach Identifies Inducers of a Mutant KRAS Selective Lethal Phenotype
Article Snippet: .. Plates were incubated for an additional 24 hours (for a total of 48 hours) under the same atmosphere and then treated with 20 μL per well of CellTiter-Glo 3D (Part#G9683, Promega Corp., Madison, WI). ..

Inhibition:

Article Title: Engineering human cell spheroids to model embryonic tissue fusion in vitro
Article Snippet: Inhibitors (or DMSO control) were diluted 1:1000 into CnT-PR-CC and added to HWJSC/HPEKp spheroids on day 0 of fusion to generate concentrations of 20 μM, 6.5 μM, and 10 μM consistent with concentrations used in the literature for in vitro screening of CH5183284[ ] and Erlotinib[ ] and for in vitro palate fusion inhibition with SB431542[ ], respectively. .. On fusion day 4, medium in each well was replaced with 100 μL per well of CellTiter Glo 3D reagent (Promega), and spheroids was incubated for 20 min on an orbital shaker at 37°C.

Proliferation Assay:

Article Title: Increase in resistance to anticancer drugs involves occludin in spheroid culture model of lung adenocarcinoma A549 cells
Article Snippet: .. In 2D and 3D culture models, the cell viability was measured using a Premix WST-1 Cell Proliferation Assay Kit (Takara, Otsu, Japan) and a CellTiter-Glo 3D Cell Viability Assay kit (Promega, Madison, WI, USA), respectively. .. Total RNA was extracted using TRI reagent (Sigma-Aldrich).

Article Title: A Novel 3-dimensional High Throughput Screening Approach Identifies Inducers of a Mutant KRAS Selective Lethal Phenotype
Article Snippet: Paragraph title: 3D Cell Culture and 3D Luminescent Proliferation Assay ... Plates were incubated for an additional 24 hours (for a total of 48 hours) under the same atmosphere and then treated with 20 μL per well of CellTiter-Glo 3D (Part#G9683, Promega Corp., Madison, WI).

Knock-Out:

Article Title: The Hsp70 co-chaperone Ydj1/HDJ2 regulates ribonucleotide reductase activity
Article Snippet: For IC50 calculations, HAP1 cells and HDJ2 Knockout were seeded in triplicate in 96-well white bottom Nunc plates in growth media at 20% confluency 1 day prior to initiation of drug treatment. .. After 72 h, cell viability was measured using Promega CellTiter-Glo cell viability assay on a Synergy H1 plate reader.

Confocal Microscopy:

Article Title: Engineering human cell spheroids to model embryonic tissue fusion in vitro
Article Snippet: Fusion was monitored each day with confocal microscopy (using 10X objective and filters for red and green wavelengths), and medium was replenished with inhibitors or EGF on day 2 of fusion. .. On fusion day 4, medium in each well was replaced with 100 μL per well of CellTiter Glo 3D reagent (Promega), and spheroids was incubated for 20 min on an orbital shaker at 37°C.

Modification:

Article Title: The Hsp70 co-chaperone Ydj1/HDJ2 regulates ribonucleotide reductase activity
Article Snippet: HAP1 cells and HDJ2 Knockout cells were obtained from Horizon Biosciences and were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM) supplemented with 10% fetal bovine serum (FBS; Invitrogen), 100 U/ml penicillin (Invitrogen) and 100 μg/ml streptomycin (Invitrogen). .. After 72 h, cell viability was measured using Promega CellTiter-Glo cell viability assay on a Synergy H1 plate reader.

Western Blot:

Article Title: Akt Regulates TNF? Synthesis Downstream of RIP1 Kinase Activation during Necroptosis
Article Snippet: Cell Viability Experiments Cells were seeded into white clear bottom 96 well plates at the density of 1×104 cells/well and treated as described for western blot experiments. .. Cell viability was determined using CellTiter-Glo Cell Viability Assay (Promega).

Cell Viability Assay:

Article Title: Near‐infrared photoimmunotherapy targeting EGFR—Shedding new light on glioblastoma treatment
Article Snippet: .. Cell response to PIT was evaluated by the CellTiter‐Glo® luminescent cell viability assay (Promega, Madison, WI) 24 or 96 h post‐irradiation in the 2D and 3D cell cultures. .. In addition, formation and growth of 3D spheroids was monitored daily by the Celigo® image cytometry system (Nexcelom Bioscience, Lawrence, MA).

Article Title: WNK1 kinase and its partners Akt, SGK1 and NBC-family Na+/HCO3− cotransporters are potential therapeutic targets for glioblastoma stem-like cells linked to Bisacodyl signaling
Article Snippet: At the end of treatments, CellTiter-Glo® luminescent cell viability assay was used according to the instructions of the manufacturer (Promega). .. For macro-spheres, cell viability assays were performed with CellTiter-Glo® 3D cell viability assay (Promega).

Recombinant:

Article Title: Engineering human cell spheroids to model embryonic tissue fusion in vitro
Article Snippet: Alternatively, spheroids were treated at day 0 with various concentrations of recombinant human EGF (BioLegend) that was reconstituted at 10 μg/mL in sterile 1% BSA in DPBS, diluted with 1% BSA in DPBS, and added to spheroids at either 2, 10, or 50 ng/mL final concentration in CnT-PR-CC with 1% antibiotic-antimycotic, with the vehicle control containing the same dilution of 1% BSA in DPBS. .. On fusion day 4, medium in each well was replaced with 100 μL per well of CellTiter Glo 3D reagent (Promega), and spheroids was incubated for 20 min on an orbital shaker at 37°C.

Viability Assay:

Article Title: The Hsp70 co-chaperone Ydj1/HDJ2 regulates ribonucleotide reductase activity
Article Snippet: .. After 72 h, cell viability was measured using Promega CellTiter-Glo cell viability assay on a Synergy H1 plate reader. .. Purification of HIS-tagged proteins from mammalian cells HEK293T cells were either un-transfected or transfected with plasmids for expression of HIS-tagged proteins using Lipofectamine 3000 (Thermo).

Article Title: WNK1 kinase and its partners Akt, SGK1 and NBC-family Na+/HCO3− cotransporters are potential therapeutic targets for glioblastoma stem-like cells linked to Bisacodyl signaling
Article Snippet: .. For macro-spheres, cell viability assays were performed with CellTiter-Glo® 3D cell viability assay (Promega). ..

Article Title: Large-scale data integration framework provides a comprehensive view on glioblastoma multiforme
Article Snippet: .. Cell proliferation was assayed 72 h after transfection using CellTiter-Glo Cell Viability assay. ..

Article Title: Akt Regulates TNF? Synthesis Downstream of RIP1 Kinase Activation during Necroptosis
Article Snippet: .. Cell viability was determined using CellTiter-Glo Cell Viability Assay (Promega). ..

Article Title: Increase in resistance to anticancer drugs involves occludin in spheroid culture model of lung adenocarcinoma A549 cells
Article Snippet: .. In 2D and 3D culture models, the cell viability was measured using a Premix WST-1 Cell Proliferation Assay Kit (Takara, Otsu, Japan) and a CellTiter-Glo 3D Cell Viability Assay kit (Promega, Madison, WI, USA), respectively. .. Total RNA was extracted using TRI reagent (Sigma-Aldrich).

Article Title: Establishment of patient-derived tumor spheroids for non-small cell lung cancer
Article Snippet: .. To demonstrate the utility of PDS as a drug screening platform, the cytotoxicity of cisplatin in PDS and H1299 tumor spheroids was investigated using CellTiter-Glo 3D Cell Viability Assay. .. The concentration of cisplatin tested ranged from 1–1000 μM with an exposure period of 72h.

Article Title: Lipid-mRNA Nanoparticle Designed to Enhance Intracellular Delivery Mediated by Shock Waves
Article Snippet: .. Cell viability was measured using CellTiter-Glo cell viability assay (Promega G7573). .. Cells were cultured in 96-well plates and, after transfection, were incubated for 24 h, the medium was removed and 100 μL of the CellTiter-Glo reagent was added to each well and mixed by gentle shaking.

Microplate Reader Luminescence Measurement:

Article Title: Engineering human cell spheroids to model embryonic tissue fusion in vitro
Article Snippet: On fusion day 4, medium in each well was replaced with 100 μL per well of CellTiter Glo 3D reagent (Promega), and spheroids was incubated for 20 min on an orbital shaker at 37°C. .. Reagent from each well was then transferred to a white opaque 96 well plate (Falcon) and read on a luminescence plate reader with 10 s integration time per well (Clariostar BMG Labtech).

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    Promega atp based cell titer glo ctg luminescent cell viability assay
    Atp Based Cell Titer Glo Ctg Luminescent Cell Viability Assay, supplied by Promega, used in various techniques. Bioz Stars score: 96/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/atp based cell titer glo ctg luminescent cell viability assay/product/Promega
    Average 96 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    atp based cell titer glo ctg luminescent cell viability assay - by Bioz Stars, 2020-04
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