Cancer cells in complete cell culture medium were seeded in 96-well plates (100 μL/well) at the optimized densities (50,000–100,000 suspension cells, 3,000–5,000 adherent cells). Suspension cells were treated 30 min after seeding, whereas adherent cells were allowed to adhere overnight and then treated. Compound treatments were prepared in complete cell culture media and 100 μL of 2X treatment-containing media were added to each well. Complete cell culture media without treatment was added in control wells, and medium alone wells were included and served as background control. The outer wells of 96-well plate were not used for treatment and were filled with 200 μL of PBS to reduce evaporation of media from inner wells. Each compound/combination was tested at nine different concentrations with three to six replicates, unless otherwise specified. For combination treatments with DT2216 and ABT199 or DT2216 and S63845, cells were treated at equimolar concentrations in two-fold serial dilutions. For combination treatment with DT2216 and chemotherapeutics, cells were treated at equimolar concentrations and three-fold serial dilutions, unless otherwise specified. The cell viability was measured after 72 h by Tetrazolium-based MTS assay. MTS reagent (2 mg/mL stock; Cat. No. G1111, Promega, Madison, WI, USA) was freshly supplemented with Phenazine methosulfate (PMS, 0.92 mg/mL stock, Cat. No. P9625, Sigma-Aldrich) in 20:1 ratio, and 20 μL of this mixture was added to each control and treatment well. The cells were incubated for four hours at 37°C and 5% CO2, and then the absorbance was recorded at 490 nm using Biotek’s Synergy Neo2 multi-mode plate reader (Biotek, Winooski, VT, USA). The average absorbance value of background control wells was subtracted from absorbance value of each vehicle-control and treatment wells and percent cell viability [(At/A0) × 100)] was determined in each treatment well, where At is the absorbance value of treatment well and A0 is the average absorbance value of control wells after background subtraction. The data were expressed as average % cell viability and fitted in non-linear regression curves using GraphPad Prism 7 (GraphPad Software, La Jolla, CA, USA).
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ABT-199
ABT-199
ABT-199 is a selective and potent inhibitor of the anti-apoptotic protein BCL-2, which plays a crucial role in the regulation of cell survival and death.
This compound has been extensively studied for its potential therapeutic applications in various malignancies, particularly in hematological cancers where BCL-2 overexpression is common.
PubCompare.ai's AI-driven protocol comparison tools can help optimize ABT-199 research by easily locating the most effective products, methods, and protocols from literature, preprints, and patents, thereby enhancing reproducibility and advancing your research.
Discover the best ABT-199 strategies to support your investigattions.
This compound has been extensively studied for its potential therapeutic applications in various malignancies, particularly in hematological cancers where BCL-2 overexpression is common.
PubCompare.ai's AI-driven protocol comparison tools can help optimize ABT-199 research by easily locating the most effective products, methods, and protocols from literature, preprints, and patents, thereby enhancing reproducibility and advancing your research.
Discover the best ABT-199 strategies to support your investigattions.
Most cited protocols related to «ABT-199»
ABT-199
Biological Assay
Cell Culture Techniques
Cells
Cell Survival
DT2216
Malignant Neoplasms
Methylphenazonium Methosulfate
Pharmacotherapy
prisma
Promega
S63845
Technique, Dilution
Tetrazolium Salts
ABT-100
ABT-199
Animals
Antibodies, Anti-Idiotypic
Asphyxia
Body Weight
Bone Marrow
Cells
Ethanol
Females
Femur
Flow Cytometry
Gamma Rays
Heterografts
Homo sapiens
Institutional Animal Care and Use Committees
Liver
Luciferases, Firefly
Luciferins
Malignant Neoplasms
Mice, Inbred NOD
Mus
Neoplasms
Patients
polyethylene glycol 400
Propylene Glycol
SCID Mice
Spleen
Tube Feeding
The materials are as follows: recombinant TNF-α (PeproTech, NJ, USA); ABT-199, SP600125, and lenalidomide (MedChemExpress, NJ, USA); TNF-α, myeloperoxidase (MPO), interleukin-1 β (IL-1β), IL-6, IL-8, and IL-18 enzyme linked immunosorbent assay (ELISA) kits (BD Biosciences, CA, USA); rabbit anti-mouse FoxO3a antibody, rabbit anti-mouse Ly6G antibody, rabbit anti-mouse JNK antibody, rabbit anti-mouse Bcl-2 antibody, rabbit anti-mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) antibody, horseradish peroxidase- (HRP-) labeled goat anti-rabbit secondary antibody (Abcam, Cambridge, UK); HRP-labeled anti-mouse secondary antibody, bicinchoninic acid (BCA) Protein Assay Kit (Beyotime Biotechnology, Shanghai, China); Naphthol AS-D Chloroacetate (Specific Esterase) Kit (Sigma, NY, USA); Diaminobenzidine DAB Kit (ZLI9019; ZSGB-BIO); Dulbecco's modified Eagle's medium (DMEM) medium, trypsin, and fetal bovine serum (FBS) (Gibco, NY, USA); FoxO3a small interfering ribonucleic acid (siRNA) (Guangzhou RiboBio Co., Ltd., Guangzhou, China); and Cell counting kit-8 (CCK-8) and Annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI) Apoptosis Assay Kit (Jiangsu KeyGEN BioTECH Corp., Ltd., Jiangsu, China).
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ABT-199
Annexin A5
Antibodies, Anti-Idiotypic
Apoptosis
bcl-2 Gene
bicinchoninic acid
Biological Assay
chloroacetate
Enzyme-Linked Immunosorbent Assay
Esterases
Fetal Bovine Serum
Fluorescein
Glyceraldehyde-3-Phosphate Dehydrogenases
Goat
Interleukin-1
interleukin 18 protein, human
isothiocyanate
Lenalidomide
Mus
Naphthols
Peroxidase
Propidium Iodide
Proteins
Rabbits
RNA
SP600125
Trypsin
Tumor Necrosis Factor-alpha
ABT-199
Alleles
Amelogenin
Cell Culture Techniques
Cell Line Authentication
Cell Lines
Cells
derivatives
Gender Identity
Genetic Profile
Genome
Homo sapiens
Lymphoma
Markers, DNA
Mus
Mycoplasma
Parent
Penicillins
Promega
Short Tandem Repeat
Streptomycin
Ficoll gradient-purified mononuclear cells from PB from newly diagnosed B-lineage ALL, from healthy volunteers, CML CD34+ cells, or BV173 or SUPB15 cells were cultured at 1 × 106 cells/ml in the presence of increasing concentrations of ABT-199 (0, 0.001, 0.01, 0.1, 1 µM) for 3 h followed by staining with 50 nM TMRE (tetramethylrhodamine ethyl ester) for 20 min at 37 °C. Separately, cells were treated with the ionophore FCCP (5 µM) (Carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone) as a positive control. MOMP of viable cells was determined by flow cytometry.
Pre-treatment of PDX cells co-cultured on bone marrow derived mesenchymal stem cells (MSCs) or of BV173 cells with dexamethasone (100 or 50 nM) and dasatinib (50 or 0.5 nM) was performed for 48 h prior to TMRE assay.
Cell culture, cloning of lentiviral constructs, apoptosis analysis, proliferation assays, immunoprecipitation, immunoblotting and statistical methods are described in detail inSupplementary Materials .
Pre-treatment of PDX cells co-cultured on bone marrow derived mesenchymal stem cells (MSCs) or of BV173 cells with dexamethasone (100 or 50 nM) and dasatinib (50 or 0.5 nM) was performed for 48 h prior to TMRE assay.
Cell culture, cloning of lentiviral constructs, apoptosis analysis, proliferation assays, immunoprecipitation, immunoblotting and statistical methods are described in detail in
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ABT-199
Apoptosis
Biological Assay
Bone Marrow Mesenchymal Stem Cells
Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
Cell Culture Techniques
Cells
Dasatinib
Dexamethasone
Ficoll
Flow Cytometry
Healthy Volunteers
Immunoprecipitation
Ionophores
mesoxalonitrile
phenylhydrazone
tetramethyl rhodamine ethyl ester
Most recents protocols related to «ABT-199»
Example 8
The ARTS Mimetic Small Molecule A4 Synergizes with BH3-Mimetic Compound
To further evaluate the therapeutic potential of the ARTS mimetic A4 molecule, combination thereof with known BH-3 mimetic compound was next examined using 293 cells stably expressing Bcl-2 cherry reporter.
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ABT-199
Apoptosis
BCL2 protein, human
Cells
Combined Modality Therapy
compound A4
inhibitors
Phenotype
Precancerous Conditions
Proteins
Prunus cerasus
Signal Transduction Pathways
Therapeutics
Transforming Growth Factor beta
Mice at advanced stages of the disease (ie, pBIC mice at 180 days with incipient splenomegaly) were randomly enrolled in the experimental groups and treated intraperitoneally either (1) once a week with 200 µg of anti-mouse CD20 (10 mg/kg, clone 5D2, isotype IgG2a, Genentech); (2) twice a week with 250 µg/dose of venetoclax (10 mg/kg, ABT-199, Chemieteck); (3) with a combination of both anti-mouse CD20 and venetoclax; or (4) twice a week with 50 µL of DMSO as the vehicle for venetoclax solubilization, corresponding to the untreated (UNT) experimental group. Treatment regimens were divided into two modalities: (1) to study overall survival, mice were treated for eight consecutive weeks and then reviewed periodically until moribund and appearance of ethical end-point criteria or (2) to study the ongoing effect of the treatment in the tumor cells and the TME, mice received four consecutive weeks of treatment (half-of-treatment duration) and then they were sacrificed and submitted to the characterization of primary tissues. Additionally, an independent cohort of pBIC mice were either untreated or treated intraperitoneally once a week with 200 µg anti-mouse PD-1 (10 mg/kg, clone RMP1-14, isotype IgG2a, BioXcell) for 2 weeks, before primary tissues were analyzed for IFN-γ expression.
ABT-199
Cells
Clone Cells
IgG2A
Immunoglobulin Isotypes
Interferon Type II
Mus
Neoplasms
Sulfoxide, Dimethyl
Tissues
Treatment Protocols
venetoclax
Freshly collected cells after cellular disaggregation of murine spleens and erythrocyte lysis using ACK buffer were incubated in RPMI media containing 10% heat-inactivated FBS and 1% penicillin/streptomycin (complete media). For drug dose-response assays, cells were incubated in triplicates for 24 hours with serial dilutions of venetoclax (0.0024–80 µM) (ABT-199, Chemieteck) or DMSO (0%–0.01% final concentration). Cell viability was assessed through flow cytometry analysis on a CytoFLEX LX flow cytometer (Beckman Coulter) after FSC/SSC/7-AAD-based exclusion of doublets and dead cells. Percentage of viable lymphoma B cells was calculated as proportion of B220+/lowCD19+GFP+ cells in a given drug concentration normalized to untreated cells at the time of treatment (24 hours after plating). The half-maximal inhibitory concentration (IC50) was determined with GraphPad Prism V.9.0.2 software using the log(inhibitor) versus normalized response equation and the least squares regression fitting method during non-linear regression modeling of the data. Percentage of apoptotic AnnexinV or active caspase 3 positive cells were calculated after DMSO baseline correction by flow cytometry as indicated above. Drug effects on T cell viability were assessed by incubating fresh splenic cell suspensions with either venetoclax IC50 (6 µM), 25 µg of anti-CD20 (clone 5D2, isotype IgG2a, Genentech) or the combination of both for 24 hours with complete media supplemented with 10% non-inactivated rat serum, followed by T-cell specific antibody staining and flow cytometry analysis as described above.
ABT-199
Apoptosis
B-Cell Lymphomas
Biological Assay
Buffers
Caspase 3
Cells
Cell Survival
Clone Cells
Erythrocytes
Flow Cytometry
IgG2A
Immunoglobulin Isotypes
Immunoglobulins
Mus
Penicillins
Pharmaceutical Preparations
prisma
Psychological Inhibition
Serum
Spleen
Streptomycin
Sulfoxide, Dimethyl
T-Lymphocyte
Technique, Dilution
venetoclax
Human MDS-derived cell lines MOLM-13, SKM-1, F-36P, and AML cell lines U937 and MV-4-11 were used in this study. For all of the experiments, the cells were maintained in RPMI-1640 supplemented with 10% fetal bovine serum and penicillin-streptomycin. All of the experiments utilized logarithmically grown cells (3–4 × 105 cells/mL). MycoAlert (Lonza, Allendale, NJ, USA) assays were performed, demonstrating that all of the cell lines were free of mycoplasma contamination. Inducible MOLM-13 BCL-2 knock-down cells were obtained as previously described [9 (link)].
We obtained 5-Aza-T-dCyd and T-dCyd from the Developmental Therapeutics Program, Division of Cancer Treatment and Diagnosis, National Cancer Institute (Rockville, MD, USA), and the Frederick National Laboratory, Frederick, MD, USA. ABT-199 was purchased from ChemieTek (Indianapolis, IN, USA). N-acetyl-L-cysteine (NAC) was purchased from Sigma-Aldrich (St. Louis, MO, USA). All of the drugs were dissolved in DMSO and reconstituted to 10 mM. The final DMSO concentrations did not exceed 0.1%. Aliquots of the drug were stored at −80 °C until use.
We obtained 5-Aza-T-dCyd and T-dCyd from the Developmental Therapeutics Program, Division of Cancer Treatment and Diagnosis, National Cancer Institute (Rockville, MD, USA), and the Frederick National Laboratory, Frederick, MD, USA. ABT-199 was purchased from ChemieTek (Indianapolis, IN, USA). N-acetyl-L-cysteine (NAC) was purchased from Sigma-Aldrich (St. Louis, MO, USA). All of the drugs were dissolved in DMSO and reconstituted to 10 mM. The final DMSO concentrations did not exceed 0.1%. Aliquots of the drug were stored at −80 °C until use.
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ABT-199
Acetylcysteine
BCL2 protein, human
Biological Assay
Cell Lines
Cells
Decitabine
Deoxycytidine
Diagnosis
Fetal Bovine Serum
Homo sapiens
Lanugo
Malignant Neoplasms
Mycoplasma
Penicillins
Pharmaceutical Preparations
Streptomycin
Sulfoxide, Dimethyl
KMS-11 cells (seeded 6 × 105 viable cells in each well, 6 replicates for each group) were treated with 5μM ABT-199, 5μM AT-101, or DMSO vehicle control in RPMI medium with 10% FBS for 24 h in a manner previously described by us.4 The Seahorse XF Cell Mito Stress Test Kit (Agilent 103015-100) was used to measuring cell mitochondrial function using the Agilent Seahorse XF96 analyzer following the manufacturer’s instructions (https://www.agilent.com/cs/library/usermanuals/public/XF_Cell_Mito_Stress_Test_Kit_User_Guide.pdf ) accessed on 4 April 2022). The assay uses the built-in injection ports on XF sensor cartridges to add modulators of respiration into cell well during the assay to reveal the key parameters of mitochondrial function. The modulators included in this assay kit are Oligomycin, Carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP), Rotenone, and Antimycin. This figure illustrates the injection sequence of these modulators and the parameters that are obtained with this assay.
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ABT-199
antimycin
Biological Assay
Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
cDNA Library
Cell Respiration
Cells
Exercise Tests
mesoxalonitrile
Mitochondrial Inheritance
Mitomycin
Oligomycins
phenylhydrazone
Physiology, Cell
Rotenone
Seahorses
Sulfoxide, Dimethyl
Top products related to «ABT-199»
Sourced in United States, Germany
ABT-199 is a small molecule inhibitor that targets the BCL-2 protein. It is primarily used for research purposes in the laboratory setting.
Sourced in United States, Germany, China
ABT-263 is a laboratory research chemical produced by Selleck Chemicals. It is a small molecule inhibitor. The core function of ABT-263 is to inhibit apoptosis regulators.
Sourced in United States, Germany
ABT-737 is a laboratory compound used in scientific research. It functions as a selective inhibitor of the Bcl-2 family of anti-apoptotic proteins. The core function of ABT-737 is to modulate apoptosis, a fundamental cellular process.
Sourced in United States
Venetoclax (ABT-199) is a selective and potent inhibitor of the anti-apoptotic protein BCL-2. It is a small molecule that binds to BCL-2, preventing it from sequestering pro-apoptotic proteins, thereby promoting apoptosis in cancer cells.
Sourced in United States, Israel
ABT-199 is a laboratory equipment product manufactured by AbbVie. It is designed for use in research and scientific applications. The core function of ABT-199 is to provide precise and reliable measurements or analysis in a laboratory setting.
Sourced in United States, Germany, Hong Kong
ABT-199 is a small-molecule inhibitor of the anti-apoptotic protein BCL-2. It functions by binding to and inhibiting the activity of BCL-2, thereby promoting apoptosis or programmed cell death in cells.
Sourced in United States, Germany, United Kingdom, China, Italy, Sao Tome and Principe, France, Macao, India, Canada, Switzerland, Japan, Australia, Spain, Poland, Belgium, Brazil, Czechia, Portugal, Austria, Denmark, Israel, Sweden, Ireland, Hungary, Mexico, Netherlands, Singapore, Indonesia, Slovakia, Cameroon, Norway, Thailand, Chile, Finland, Malaysia, Latvia, New Zealand, Hong Kong, Pakistan, Uruguay, Bangladesh
DMSO is a versatile organic solvent commonly used in laboratory settings. It has a high boiling point, low viscosity, and the ability to dissolve a wide range of polar and non-polar compounds. DMSO's core function is as a solvent, allowing for the effective dissolution and handling of various chemical substances during research and experimentation.
Sourced in United States
ABT-199 is a selective and potent inhibitor of the anti-apoptotic protein BCL-2. It is commonly used in research applications to study the role of BCL-2 in cellular processes and disease models.
Sourced in United States
ABT-199 is a small molecule inhibitor that selectively targets the anti-apoptotic protein BCL-2. It is designed for use in research applications.
Sourced in United States, Germany, United Kingdom
S63845 is a laboratory instrument designed for the purpose of analytical measurement. The core function of this product is to provide precise and reliable data analysis capabilities within a controlled laboratory environment.
More about "ABT-199"
ABT-199, also known as Venetoclax, is a highly selective and potent inhibitor of the anti-apoptotic protein BCL-2, which plays a crucial role in regulating cell survival and death.
This compound has been extensively studied for its potential therapeutic applications, particularly in hematological cancers where BCL-2 overexpression is common.
PubCompare.ai's AI-driven protocol comparison tools can help optimize your ABT-199 research by easily locating the most effective products, methods, and protocols from literature, preprints, and patents.
This can enhance reproducibility and advance your investigations into this promising compound.
Aside from ABT-199, related BCL-2 inhibitors like ABT-263 (Navitoclax) and ABT-737 have also been studied for their anti-cancer properties.
Similarly, the small molecule inhibitor S63845, which targets the MCL-1 protein, has shown synergistic effects when combined with ABT-199.
When working with ABT-199, it's important to use appropriate vehicle controls, such as DMSO, to ensure accurate and reliable results.
PubCompare.ai's tools can help you identify the best protocols and methods for your ABT-199 research, driving your investigations forward and ultimately supporting the development of new therapies.
This compound has been extensively studied for its potential therapeutic applications, particularly in hematological cancers where BCL-2 overexpression is common.
PubCompare.ai's AI-driven protocol comparison tools can help optimize your ABT-199 research by easily locating the most effective products, methods, and protocols from literature, preprints, and patents.
This can enhance reproducibility and advance your investigations into this promising compound.
Aside from ABT-199, related BCL-2 inhibitors like ABT-263 (Navitoclax) and ABT-737 have also been studied for their anti-cancer properties.
Similarly, the small molecule inhibitor S63845, which targets the MCL-1 protein, has shown synergistic effects when combined with ABT-199.
When working with ABT-199, it's important to use appropriate vehicle controls, such as DMSO, to ensure accurate and reliable results.
PubCompare.ai's tools can help you identify the best protocols and methods for your ABT-199 research, driving your investigations forward and ultimately supporting the development of new therapies.