Curcumin, AO, MDC, NAC, z-VAD, CHX, and DCF-DA were purchased from Sigma (St. Louis, MO, USA). The class III PI3K inhibitor 3-MA was obtained from Calbiochem (La Jolla, CA, USA). The CellTiter 96 AQueous One Solution Proliferation Assay System was purchased from Promega (Madison, WI, USA). Antibodies against PARP, caspase-3, p62, Beclin-1, CHOP, phospho-eIF2α (Ser51), IRE1-α, Bip/GRP78 and Nrf2 were purchased from Cell Signaling Technology (Beverly, MA, USA). Antibodies against LC3 (Sigma) were also used. The anti-β-actin, anti-ubiquitin, goat anti-rabbit and anti-mouse IgG antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
Celltiter 96 aqueous one solution proliferation assay system
Manufactured by Promega
Sourced in United States
The CellTiter 96 AQueous One Solution Proliferation Assay System is a colorimetric method for determining the number of viable cells in proliferation assays. The assay is based on the use of a tetrazolium compound that is reduced by metabolically active cells, producing a colored formazan product that can be measured spectrophotometrically.
Automatically generated - may contain errors
Lab products found in correlation
Fitc annexin 5 apoptosis detection kit 2, by BD (2 mentions)
β actin, by Santa Cruz Biotechnology (2 mentions)
Bip grp78, by Cell Signaling Technology (1 mentions)
Antibodies against lc3, by Merck Group (1 mentions)
Anti β actin, by Santa Cruz Biotechnology (1 mentions)
Beclin 1, by Cell Signaling Technology (1 mentions)
Caspase 3, by Cell Signaling Technology (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Phospho eif2α ser51, by Cell Signaling Technology (1 mentions)
Z vad, by Merck Group (1 mentions)
Anti ubiquitin, by Santa Cruz Biotechnology (1 mentions)
Ire1α, by Cell Signaling Technology (1 mentions)
Anti mouse igg, by Santa Cruz Biotechnology (1 mentions)
Goat anti rabbit, by Santa Cruz Biotechnology (1 mentions)
N acetyl cysteine (nac), by Merck Group (1 mentions)
Curcumin, by Merck Group (1 mentions)
Molecular weight standards, by Thermo Fisher Scientific (1 mentions)
Ab229935, by Abcam (1 mentions)
E cadherin 3195, by Cell Signaling Technology (1 mentions)
Sc 8396, by Santa Cruz Biotechnology (1 mentions)
9 protocols using celltiter 96 aqueous one solution proliferation assay system
1
Curcumin Modulates Cellular Stress Responses
2
Comprehensive Materials and Reagents for Cell Assays
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API (A3145) and dimethyl sulfoxide (DMSO) were purchased from Sigma-Aldrich (St. Louis, MO). Fetal bovine serum (FBS), antibiotics, molecular weight standards, trypsin-EDTA, and all medium additives were obtained from Life Technologies (Gaithersburg, MD). The CellTiter 96 AQueous One Solution Proliferation Assay System was purchased from Promega (Madison, WI). An enhanced chemiluminescence (ECL) kit was purchased from Amersham (Arlington Heights, IL). Antibodies specific for fibronectin (ab2413) and SPOCK1 (ab229935) were obtained from Abcam (Cambridge, MA). Antibodies specific for cleaved-PARP (#9541), phospho-Ak (ser473, #9271), Akt (#4691), Snail (#3895), Slug (#9585), Twist (#46702), and E-cadherin (#3195) were obtained from Cell Signaling Technology (Danvers, MA). Antibodies specific for vimentin (550513) and N-cadherin (610920) were purchased from BD Biosciences (San Jose, CA). Antibodies specific for cyclin D1 (sc-8396), cyclin E (sc-377,100), β-actin (sc-47,778), and goat anti-rabbit (sc-2004) and anti-mouse (sc-2005) immunoglobulin G (IgG) antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Polyvinylidene fluoride (PVDF) membranes for Western blotting were purchased from Bio-Rad (Hercules, CA). Unless otherwise specified, other chemicals used in this study were purchased from Sigma Chemical (St. Louis, MO).
3
CXCR4 Antibody-Conjugated GNPs Cytotoxicity
4
Cell Viability Assay of Compound PM
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Tumor cells (1×104) in 100 μl of tissue culture medium were seeded into each well of a 96-well plate. After 24-h incubation to allow cells to adhere, cells were treated with PM at concentrations ranging from 0 to 5 μM. Cultures were incubated for additional 72 h and cell viability was then determined by the colorimetric MTS assay using CellTiter 96 AQueous One Solution Proliferation Assay System from Promega. This assay measures the bioreduction of the tetrazolium compound MTS by intracellular dehydrogenases in the presence of electron-coupling reagent phenazine methosulfate. MTS and phenazine methosulfate were added to the culture wells, and cultures were incubated for 2 h at 37°C. The absorbance, which is directly proportional to the number of viable cells in the cultures, was measured at 490 nm using a microplate reader.
5
Curcumin Cytotoxicity Assay in Cells
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Cells (1 × 104) were seeded into each well of a 96-well plate in 100 μL of tissue culture medium. After 24 h of incubation to allow the cells to adhere, the cells were treated with curcumin and incubated for an additional 48 h. Cell viability was then determined by performing the colorimetric MTS assay using the CellTiter 96 AQueous One Solution Proliferation Assay System (Promega). This assay measures the bioreduction of the tetrazolium compound MTS by intracellular dehydrogenases in the presence of the electron-coupling reagent phenazine methosulfate. MTS and phenazine methosulfate were added to the culture wells, and the mixture was incubated for 2 h at 37 °C. The absorbance, which is directly proportional to the number of viable cells in culture, was measured at 490 nm using a microplate reader. The percentage of cytotoxicity was calculated based on the loss of viable cells in the cultures.
6
Assessing Cell Viability via MTS Assay
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Tumor cells (1×104) in 100 μl of tissue culture medium were seeded into each well of a 96-well plate. After 24-h incubation to allow cells to adhere, cells were treated with PM at concentrations ranging from 0 to 5 μM. Cultures were incubated for additional 72 h and cell viability was then determined by the colorimetric MTS assay using CellTiter 96 AQueous One Solution Proliferation Assay System from Promega. This assay measures the bioreduction of the tetrazolium compound MTS by intracellular dehydrogenases in the presence of electron-coupling reagent phenazine methosulfate. MTS and phenazine methosulfate were added to the culture wells, and cultures were incubated for 2 h at 37°C. The absorbance, which is directly proportional to the number of viable cells in the cultures, was measured at 490 nm using a microplate reader.
7
Cell Viability, Cytotoxicity, and Apoptosis Assays
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The cell viability was detected by the colorimetric MTS assay using the CellTiter 96 Aqueous One Solution proliferation assay system (Promega, Madison, WI). LDH, a stable cytosolic enzyme that is released upon cell lysis was measured by the CytoTox 96 non-radioactive cytotoxicity assay (Promega, Madison, WI). Active Caspase 3 was assay with the caspase 3/7-GLO assay (Promega, Madison, WI).
8
Telomerase and Apoptosis Assays for PM
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PM was purchased from Sigma Chemicals (St. Louis, MO, USA). Antibodies against PARP-1, Akt, p-Akt (Ser473), Sp1, c-Myc, NF-κB and β-actin were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Anti-hTERT and p-TERT antibodies were obtained from Abcam, Inc. (Cambridge, MA, USA). The CellTiter 96® AQueous One Solution Proliferation Assay System was purchased from Promega (Madison, WI, USA). The Annexin V-FITC Apoptosis Detection Kit II was obtained from BD Biosciences Pharmingen (San Diego, CA, USA) and the TRAPeze Telomerase Detection kit was purchased from Millipore (Temecula, CA, USA).
A stock solution (100 mM) of PM was prepared in dimethyl-sulfoxide (DMSO) and test concentrations were prepared by diluting the stock solution in tissue culture medium.
A stock solution (100 mM) of PM was prepared in dimethyl-sulfoxide (DMSO) and test concentrations were prepared by diluting the stock solution in tissue culture medium.
9
CDDO-Me Inhibits Akt and mTOR Signaling
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CDDO-Me was obtained from the National Cancer Institute, Bethesda, MD, USA through the Rapid Access to Intervention Development Program. Antibodies against p-Akt (ser473), p-mTOR (Ser2448), PARP-1 and β-actin were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). CellTiter 96 AQueous One Solution Proliferation assay system was from Promega (Madison, WI, USA) and Annexin V-FITC apoptosis detection kit II was obtained from BD Pharmingen (San Diego, CA, USA).
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