Ic50 calculator

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The IC50 calculator is a software tool that determines the half-maximal inhibitory concentration (IC50) of a compound. It analyzes experimental data and calculates the concentration at which a compound inhibits a specific biological or biochemical process by 50%. The IC50 calculator provides a quantitative measure of a compound's potency, which is a key metric in drug discovery and development.

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Lab products found in correlation

Microplate reader, by Tecan (3 mentions) M5655, by Merck Group (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions) Microplate reader, by Bio-Rad (1 mentions) Prism 5, by GraphPad (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Penicillin, by American Type Culture Collection (1 mentions) Streptomycin, by American Type Culture Collection (1 mentions) 3t3 l1 cells, by American Type Culture Collection (1 mentions) Cell counting kit 8 (cck8), by Dojindo Laboratories (1 mentions) Gen5 spectrophotometer, by Agilent Technologies (1 mentions) Dinaciclib, by Selleck Chemicals (1 mentions) Abemaciclib, by Selleck Chemicals (1 mentions) Opsys mr, by Dynex (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)

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Quest Graph™ IC50 Calculator (47 citations) MLA—Quest Graph™ IC50 Calculator (6 citations) IC50 Calculator software (2 citations)

15 protocols using ic50 calculator

Statistical Analysis of Drug Sensitivity

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The significance of differences in growth rate and sensitivity to drugs was assessed through the two-tailed unpaired Student’s t-test or t-test with Welch’s correction, according to assumptions of the tests and the variance between the compared groups. The used test was reported in each figure legend.
Calculations of the IC₅₀ (half maximal inhibitory concentration) of the drugs used in this paper were based on the interpolation of the growth percentages with a sigmoid dose-response curve by Prism 5 software (GraphPad software, La Jolla, CA, USA) and IC₅₀ Calculator | AAT Bioquest (IC₅₀ Calculator | AAT Bioquest). The significance of differences in IC₅₀ between different cell lines and different drugs was assessed by calculating the IC₅₀value for each replicate and comparing the values of each group through the Student’s t-test. Statistical analyses were performed through Prism 5 software.

Ruzzi F., Angelicola S., Landuzzi L., Nironi E., Semprini M.S., Scalambra L., Altimari A., Gruppioni E., Fiorentino M., Giunchi F., Ferracin M., Astolfi A., Indio V., Ardizzoni A., Gelsomino F., Nanni P., Lollini P.L, & Palladini A. (2022). ADK-VR2, a cell line derived from a treatment-naïve patient with SDC4-ROS1 fusion-positive primarily crizotinib-resistant NSCLC: a novel preclinical model for new drug development of ROS1-rearranged NSCLC. Translational Lung Cancer Research, 11(11), 2216-2229.

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Antiviral Compound Evaluation: Selectivity Index

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The 50% effective concentrations (EC50), defined as a compound’s concentration inhibiting viral replication by 50%, was determined using a four- or three-parameter logistic regression (AAT Bioquest IC50 Calculator) to fit the dose–response curves. The cytotoxic concentration 50 (CC50), defined as a compound’s concentration reducing cell viability by 50%, was determined for each compound and cell type by the same method (AAT Bioquest IC50 Calculator). When our measurements did not reach 50% toxicity or 50% inhibition by studied concentrations, we used the estimated EC50 or CC50 value for further calculation of Selectivity Index (indicated as > 200 in Tables 2, 3). The Selectivity Index (SI) was calculated as previously reported^{21 (link)} by dividing the CC50 by the EC50 of every compound for each virus tested. A low SI (< 1) indicates a poor candidate for further development in therapeutical antiviral strategies. A high selectivity index (SI > 1) reflects an important range of concentrations that show an effective antiviral effect with low toxicity.

Pasquereau S., Galais M., Bellefroid M., Pachón Angona I., Morot-Bizot S., Ismaili L., Van Lint C, & Herbein G. (2022). Ferulic acid derivatives block coronaviruses HCoV-229E and SARS-CoV-2 replication in vitro. Scientific Reports, 12, 20309.

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Assessing A549 Cell Viability Using MTT Assay

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A549 cell viability was assessed using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) tetrazolium reduction assay (M5655, Sigma Aldrich, Milan, Italy), as described in [58 (link)]. Briefly, A549 cells were treated for 48 h in complete medium with TGFβ (10 ng/mL). Next, cells (2.5 × 10³) were plated into 96-multiwell plates in 100 µL of complete medium. After adhesion, the compounds were added to the medium at different concentrations. After 24 h exposure, the medium was replaced with 100 µL MTT reagent in each well, and plates were incubated at 37 °C. After 2 h, MTT was removed, and the blue MTT–formazan product was solubilized with dimethyl sulfoxide (DMSO, Sigma Aldrich). The absorbance of the formazan solution was read at 595 nm using the microplate reader (Bio-Rad, Milan, Italy). Inhibition of cell proliferation was calculated by the IC₅₀ calculator (AAT Bioquest, Inc., Pleasanton, CA, USA, https://www.aatbio.com/ (accessed on 1 July 2022)).

Andreucci E., Bugatti K., Peppicelli S., Ruzzolini J., Lulli M., Calorini L., Battistini L., Zanardi F., Sartori A, & Bianchini F. (2023). Nintedanib-αVβ6 Integrin Ligand Conjugates Reduce TGFβ-Induced EMT in Human Non-Small Cell Lung Cancer. International Journal of Molecular Sciences, 24(2), 1475.

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RO-3306 Inhibits OVCAR5 and SKOV3 Proliferation

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The OVCAR5 and SKOV3 cells were cultured overnight in 96-well plates with a concentration of 4000 cells/well. The cells were then treated with various concentrations of RO-3306 for 72 h. A total of 5 µL of 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium Bromide (MTT, 5 mg/mL) were added into each well, followed by incubation for 1 to 1.5 h. MTT absorbance was detected using a microplate reader (Tecan, Morrisville, NC, USA) at a wavelength of 562 nm after mixing 100 µL dimethyl sulfoxide (DMSO) into each well. The effect of RO-3306 on cell proliferation was calculated as a percentage of control, and the IC50 was calculated using the AAT Bioquest IC50 calculator (Pleasanton, CA, USA). Each experiment was performed at least three times for consistency.

Huang Y., Fan Y., Zhao Z., Zhang X., Tucker K., Staley A., Suo H., Sun W., Shen X., Deng B., Pierce S.R., West L., Yin Y., Emanuele M.J., Zhou C, & Bae-Jump V. (2023). Inhibition of CDK1 by RO-3306 Exhibits Anti-Tumorigenic Effects in Ovarian Cancer Cells and a Transgenic Mouse Model of Ovarian Cancer. International Journal of Molecular Sciences, 24(15), 12375.

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Evaluation of antivirals against SARS-CoV-2

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The prepared cells were incubated with TS at seven different concentrations as follows. The concentrations used for E4, were 5.5, 2.75, 1.37, 0.68, 0.34, 0.17 and 0.08 µg. The concentrations used for Remdesivir were 10, 3, 1, 0.3, 0.1, 0.03 and 0.01 µM. The control cells were incubated with 0.5% DMSO.
The cells were infected with SARS-CoV2 at a MOI of 0.01 and after 48 h, viral RNA was extracted from 100 µl culture supernatant and subjected to qRT-PCR. The Ct values for N and E gene sequence were recorded and the inhibition of virus replication was determined based on the change in the Ct value in TS-treated cells compared to the control. IC₅₀ values were determined using AAT Bioquest IC₅₀ calculator.

Latha D., Hrishikesh D., Shiban G., Chandrashekar C, & Bharath B. (2022). In silico, in vitro screening of plant extracts for anti-SARS-CoV-2 activity and evaluation of their acute and sub-acute toxicity. Phytomedicine plus, 2(2), 100233.

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Antiviral Efficacy of AND-2-HyP-β-CYD Complex

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The antiviral testing of solid complex was carried out using qRT-PCR assay method [25 ]. Briefly, 1 × 10e⁴ Vero E6 cells were suspended per well and incubated at 37 °C for 8 h. Further, Vero E6 cells were incubated with 0.02, 0.05, 0.11, 0.22, 0.45, 0.9, and 1.8 μg.mL⁻¹ concentration of AND-2-HyP-β-CYD complex. On the other hand, 1% ethanol only was incubated with control cells. The Vero E6 cells were infected with SARS-CoV-2 at MOI (Multiplicity of Infection) of 0.01. Following this, viral RNA was isolated from 100 μl culture supernatant after 48 h and this was subjected to qRT-PCR (in duplicates). In this way, Ct (Cycle Threshold Value) values for N and E gene sequence were determined. Suppression of virus replication was observed on the fold change in the Ct value in AND-2-HyP-β-CYD complex treated cells compared to the control. Remdesivir was employed as a standard for virus inhibition. IC₅₀ values were determined using AAT Bioquest IC50 calculator.

Singh S.C., Khatri D.K., Singh K., Kanchupalli V.K., Madan J., Singh S.B, & Singh H. (2021). Molecular encapsulation of andrographolide in 2-hydroxypropyl-β-cyclodextrin cavity: synthesis, characterization, pharmacokinetic and in vitro antiviral activity analysis against SARS-CoV-2. Heliyon, 7(8), e07741.

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Chemotherapeutic Sensitivity Comparison

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The half maximal inhibitory concentration (IC50) for each cell line histology classification (iUC versus PCA) following exposure to each chemotherapeutic agent (carboplatin, mitoxantrone, vinblastine, and docetaxel) was extracted using AAT Bioquest IC50 calculator [28 ] 4 parameter fitting. IC50 values for curves failing the 4 parameter fitting (n = 12) were generated with 2 parameter (point-to-point, n = 2) or 3 parameter (n = 10) fitting. Given that normality was not met using the Shapiro-Wilk test, Wilcoxon rank-sum testing was utilized to determine if IC50 differences existed between iUC and PCA cell lines following 72 hour exposure to each chemotherapeutic agent. RStudio (RStudio 2021, Boston, MA) was used for statistical testing, and significance was defined as p < 0.05.

Berry M.R., Fadl-Alla B.A., Samuelson J., Rosol T.J, & Fan T.M. (2022). Investigating PSMA differential expression in canine uroepithelial carcinomas to aid disease-based stratification and guide therapeutic selection. BMC Veterinary Research, 18, 441.

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Cell Proliferation Assay of IPAT and Carboplatin

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Cell proliferation was assessed in ARK-1 and SPEC-2 by MTT assay. Briefly, cells (4000–6000/well) were seeded in 96-well plates and cultured at 37 °C in 5% CO2 overnight and then treated with the indicated doses of IPAT and carboplatin as single agents or in combination for 72 h. 100 ul of MTT solution (5 mg/ml) was added to each well and incubated for another 2 h. The supernatants were aspirated and 100 µl of DOMSO was added to each well to lyse the formazan crystal. Optical density was measured at a wavelength of 570 nm with a Tecan microplate reader (Morrisville, NC). The effect of IPAT and carboplatin on proliferation was assessed as the percentage of inhibition compared to untreated cells. IC50 calculator (AAT Bioquest; Sunnyvale, CA) was used to detect IC50 for IPAT and carboplatin. Bliss Independence model was used to determine whether drug effects are additive (CI = 1), synergistic (CI < 1) or antagonistic (CI > 1) [26 (link)]. Results represent the median of three separate experiments, each performed in quadruplicate.

Burkett W.C., Zhao Z., Newton M.A., Sun W., Deng B., Secord A.A., Zhou C, & Bae-Jump V. (2023). Ipatasertib, an oral AKT inhibitor, in combination with carboplatin exhibits anti-proliferative effects in uterine serous carcinoma. Annals of Medicine, 55(1), 603-614.

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Evaluating HIV-1 RT Inhibition Potency

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The half-maximal inhibitory concentration (IC₅₀) of the compound is defined as the amount of compound required to block HIV-1 RT activity by 50%. The best-fit curves were plotted using the relative amount of the amplified GAPDH as calculated in the previous step (using ΔCt method). Curve fitting was applied using Hill equation [49 (link)] and IC₅₀ was obtained directly from the calculated best fit curve parameters using AAT BioQuest IC₅₀ calculator [50 ].

Chan K.F., Su C.T., Krah A., Phua S.X., Yeo J.Y., Ling W.L., Bond P.J, & Gan S.K. (2020). An Alternative HIV-1 Non-Nucleoside Reverse Transcriptase Inhibition Mechanism: Targeting the p51 Subunit. Molecules, 25(24), 5902.

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Cell Viability Assay for ASP and PTX

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The MES and MES-TP cells were cultured in 96-well plates with 4000 cells/well. After 24 h of incubation, the cells were treated with ASP, PTX or the combination for 72 h, and then 5 ul MTT (5 mg/ml) was added to each well for 1 h at 37 °C. The cells were then lysed with 100 ul DMSO/well, and the absorbance at 575 nm was measured using a microplate reader (Tecan, Durham, NC). The IC50 value for ASP and PTX was then calculated by the IC50 Calculator (AAT Bioquest, Sunnyvale, CA).

Zhang X., Wang J., Fan Y., Zhao Z., Paraghamian S.E., Hawkins G.M., Buckingham L., O’Donnell J., Hao T., Suo H., Yin Y., Sun W., Kong W., Sun D., Zhao L., Zhou C, & Bae-Jump V.L. (2022). Asparagus officinalis combined with paclitaxel exhibited synergistic anti-tumor activity in paclitaxel-sensitive and -resistant ovarian cancer cells. Journal of Cancer Research and Clinical Oncology, 149(7), 3871-3883.

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