Q vd oph

Manufactured by MedChemExpress

Sourced in United States

Q-VD-OPh is a laboratory reagent that functions as a pan-caspase inhibitor, capable of blocking the activity of multiple caspase enzymes. It is commonly used in research applications to study apoptosis and cell death pathways.

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Lab products found in correlation

Superscript 3 first strand synthesis system, by Thermo Fisher Scientific (2 mentions) Trizol reagent, by Thermo Fisher Scientific (2 mentions) Blasticidin, by Thermo Fisher Scientific (1 mentions) S63845, by Selleck Chemicals (1 mentions) Hs27a, by American Type Culture Collection (1 mentions) Venetoclax abt 199, by MedChemExpress (1 mentions) Isopropyl β d 1 thiogalactopyranoside (iptg), by MedChemExpress (1 mentions) Silmitasertib cx 4945, by Selleck Chemicals (1 mentions) Hek293t 17, by American Type Culture Collection (1 mentions) Mk 2206, by Selleck Chemicals (1 mentions) Puromycin, by Merck Group (1 mentions) Cycloheximide (chx), by Merck Group (1 mentions) Granta 519, by Leibniz Institute DSMZ (1 mentions) Jeko 1, by Leibniz Institute DSMZ (1 mentions) Maver 1, by Leibniz Institute DSMZ (1 mentions) Cisplatin, by Abcam (1 mentions) S63845, by MedChemExpress (1 mentions) Abt 199, by Active Motif (1 mentions) Wehi 539, by MedChemExpress (1 mentions) Venetoclax, by Selleck Chemicals (1 mentions)

9 protocols using q vd oph

Co-culture Assay for MCL Cell Lines

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The human MCL cell lines Granta-519, Jeko-1, Maver-1, Mino, and Z138 were obtained from DSMZ (Braunschweig, Germany) and cultured in Iscove’s modified Dulbecco’s medium (IMDM; Invitrogen Life Technologies, Carlsbad, CA, USA). The stromal cell line HS-27a and the kidney cell line HEK-293T/17 were obtained from ATCC (Manassas, VA, USA), and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Invitrogen).
For co-culture assays, bone-marrow stromal cells were seeded in 96-well plates 4 hours (h) prior to the addition of MCL cells, to allow cell attachment. MCL cells were pre-incubated for 2 h with stromal cells, followed by a 3-day co-culture in the presence of the indicated drug concentrations. Cell viability and specific cell death were measured by flow cytometry.
The following small-molecule inhibitors were used: venetoclax/ABT-199, Q-VD-OPh, IPTG (MedChemExpress, Princeton, NJ, USA), silmitasertib/CX-4945, S63845, MK2206 (Selleckchem, Houston, TX, USA), cycloheximide, puromycin (Sigma-Aldrich, St. Louis, MO, USA), and blasticidin (Thermo Fisher Scientific, Waltham, MA, USA).

Thus Y.J., de Rooij M.F., Swier N., Beijersbergen R.L., Guikema J.E., Kersten M.J., Eldering E., Pals S.T., Kater A.P, & Spaargaren M. (2022). Inhibition of casein kinase 2 sensitizes mantle cell lymphoma to venetoclax through MCL-1 downregulation. Haematologica, 108(3), 797-810.

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Chemotherapeutic Drug Reconstitution Protocol

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Cisplatin was purchased from Abcam (Cambridge, UK) and reconstituted in 0.9% w/v NaCl. All other agents were reconstituted in DMSO. ABT-199 was purchased from Active Biochem (Hong Kong, China) and S-63845, WEHI-539 and Q-VD-OPh from Medchemexpress (NJ, US).

Fitzgerald M.C., O’Halloran P.J., Kerrane S.A., Ní Chonghaile T., Connolly N.M, & Murphy B.M. (2023). The identification of BCL-XL and MCL-1 as key anti-apoptotic proteins in medulloblastoma that mediate distinct roles in chemotherapy resistance. Cell Death & Disease, 14(10), 705.

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Venetoclax-induced Apoptosis Pathway

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We obtained Venetoclax (also called ABT-199) from Selleckchem (Houston, TX). We purchased Pan-caspase inhibitor Q-VD-OPh from MedChemExpress (Monmouth Junction, NJ) and added to cells at 20 μM. We obtained Sodium butyrate from Wako (FUJIFILM Wako Pure Chemical, Osaka, Japan) and made use of it in this experiment as NaB. We dissolved all reagents into a solvent Dimethyl sulfoxide (DMSO).

Kawakatsu R., Tadagaki K., Yamasaki K, & Yoshida T. (2024). Venetoclax efficacy on acute myeloid leukemia is enhanced by the combination with butyrate. Scientific Reports, 14, 4975.

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Apoptosis Induction Pathway Modulation

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Artesunate, MG132, EB, and AO were purchased from Sigma-Aldrich Inc. Venetoclax (ABT-199) was purchased from Selleck Chemical. Cytarabine and Q-VD-OPh were purchased from MedChemExpress. Antibodies to PARP and caspase-8 were obtained from BD Biosciences. Antibodies to Bcl-2 (C-2), Actin (C-2), Mcl-1 (S-19), Mcl-1 (G-7), Bax (6A7) and Chk1 (G-4) were obtained from Santa Cruz Biotechnology, Inc. Antibodies to Mcl-1 (D35A5), Bim (C34C5), Bak (D4E4), Bax poly, Noxa (D8L7U), Cleaved Caspase-3 (Asp175), Phospho-Chk1 (Ser345) (133D3), and Phospho-Histone H2A.X (Ser139) (20E3) were obtained from Cell Signaling Technology, Inc. Antibodies to Noxa were obtained from Abcam, Inc. Bak (Ab-1) was obtained from Merck Millipore. NOXA(sc-37305), BIM(sc-29802), MCL1(sc-35877)siRNA, and a control siRNA were purchased from Santa Cruz Biotechnology, Inc. NOXA(s10708), BIM(s195011), MCL1(s8583)siRNA was purchased from Thermo Fisher Scientific.

Zhang J., Wang Y., Yin C., Gong P., Zhang Z., Zhao L., Waxman S, & Jing Y. (2022). Artesunate improves venetoclax plus cytarabine AML cell targeting by regulating the Noxa/Bim/Mcl-1/p-Chk1 axis. Cell Death & Disease, 13(4), 379.

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Culturing and Treating Cancer Cell Lines

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Cancer cell lines used in this study were grown in Dulbecco’s Modified Eagle’s Medium (DMEM; Genesee Scientific, San Diego, CA, #25-500) (786-O, RCC4, UM-RC2, MDA-MB-231, MCF-7), RPMI 1640 (Thermo Fisher Scientific, Waltham, MA, #A1049101) (TK-10, T-47D), McCoy’s 5A (Genesee Scientific, #25-518) (Caki-1, Caki-2), or Eagle’s Minimum Essential Medium (EMEM; Sigma-Aldrich, St. Louis, MO, #M4655) (MDCK) supplemented with 10% Fetal Bovine Serum (FBS; Genesee Scientific, #25-514) and 1% Penicillin/Streptomycin (P/S; Genesee Scientific, #25-512) in 5% CO₂ at 37°C. RPTEC cells were grown in DMEM/F12 medium with 36 ng/mL hydrocortisone (Thermo Fisher Scientific, #SH30261.01, #AC352450010), 5 μg/mL insulin, 5 μg/mL apo-transferrin, 5 ng/mL sodium selenite (Sigma-Aldrich, #I0516-5ML, #T1147-100 MG, #S5261-10 G), 50 ng/mL human EGF (PeproTech, Cranbury, NJ #AF-100-15), 1% P/S, and 100 μg/mL G418 (Selleck Chemicals, Houston, TX, #S3028). Dinaciclib and Q-VD-OPh (MedChem Express, LLC, Monmouth Junction, NJ, #HY-10492, #HY-12305) were diluted in Dimethyl Sulfoxide (DMSO) and serially diluted for each experiment. ABT-263 (BioVision Inc., Milpitas, CA, #2467-5), Staurosporine (BioVision Inc., #1048-01) were diluted in DMSO and used at concentrations indicated in the manuscript.

Nelson L.J., Castro K.E., Xu B., Li J., Dinh N.B., Thompson J.M., Woytash J., Kipp K.R, & Razorenova O.V. (2022). Synthetic lethality of cyclin-dependent kinase inhibitor Dinaciclib with VHL-deficiency allows for selective targeting of clear cell renal cell carcinoma. Cell Cycle, 21(10), 1103-1119.

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Artemisinins Induce Fibroblast Apoptosis

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ART was provided by Li Shizhen Pharmaceutical Co., Ltd. (Hubei, China). Fibroblasts were cultured in 6-well plates, 96-well plates, or 10-cm-diameter culture dishes. GSK2606414, a PERK signaling pathway inhibitor, was purchased from Medchem Express Co., Ltd. (Shanghai, China). Q-VD-Oph, a caspase signaling pathway inhibitor, was purchased from Medchem Express Co., Ltd. (Shanghai, China). When the cells were grown to nearly 80%, they were rinsed with phosphate buffer saline (PBS) for three times. Human fibroblasts were treated with ART. The control group was incubated with PBS. Subsequently, experimental and control cells were collected for subsequent cell experiments.

Chen H., Tao J., Wang J, & Yan L. (2019). Artesunate prevents knee intraarticular adhesion via PRKR-like ER kinase (PERK) signal pathway. Journal of Orthopaedic Surgery and Research, 14, 448.

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Quantitative RT-PCR Analysis of Apoptosis Genes

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To obtain intact cells for qRT-PCR, cells were incubated with 25 μM of the broad-spectrum caspase inhibitor Q-VD-OPh (MedChem Express # HY-12305) for 15 min prior to treatment with 5’azacytidine or Nutlin-3a. After 24 h, cell pellets were collected and resuspended in 0.5 mL TRIzol reagent (Thermo Fisher #15596026). RNA was isolated according to the manufacturer’s instructions. cDNA was prepared using the Superscript III First Strand Synthesis System (Thermo Fisher #18080051) according to the manufacturer’s instructions. qRT-PCR reactions were performed using TaqMan Fast Advanced Master Mix (Thermo Fisher, #4444963) according to the manufacturer’s instructions. TaqMan probes to detect mouse Puma/Bbc3 (Mm00519268_m1), Noxa/Pmaip1 (Mm00451763_m1), Bax (Mm00432050_m1) and Hmbs (Mm01143545_m1) were used (Thermo Fisher). Reactions were run on a Quantstudio 12 K Flex Real-Time PCR System (Thermo Fisher). Cycle threshold (Ct) values for each gene were normalised to the housekeeping control gene (Hmbs) Ct value for that sample and all data are presented relative to the DMSO-treated parental cell line. All qRT-PCR data are presented as mean ± standard deviation of two independent experiments.

Diepstraten S.T., Young S., La Marca J.E., Wang Z., Kluck R.M., Strasser A, & Kelly G.L. (2023). Lymphoma cells lacking pro-apoptotic BAX are highly resistant to BH3-mimetics targeting pro-survival MCL-1 but retain sensitivity to conventional DNA-damaging drugs. Cell Death and Differentiation, 30(4), 1005-1017.

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Cell Proliferation and Apoptosis Assay Protocol

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To measure cell proliferation, cells were plated in 96- or 384-well plates (3000 or 1250 cells/well, 3–6 wells/sample) with drugs and incubated for 48 hours at 37°C in 5% CO₂. After incubation, relative numbers of viable cells were measured using a tetrazolium-based colorimetric assay (CellTiter Aqueous One Solution Cell Proliferation Assay, Promega). Inhibitor combination screening was carried out in pre-treated 384 well plates with inhibitors prepared in a 7-point concentration series ranging from 10 μM to 0.014 μM for each drug (23 (link)).
To measure cell apoptosis, cells (in duplicates) were resuspended in 100 μL of Annexin V binding buffer containing 1 μL of Annexin V-PE, 1 μL of 7-aminoactinomycin D (7-AAD) (Southern Biotech) and 1 μL of CD19-mAbs (BD Bioscience) followed by flow cytometry on FACSAria (BD Biosciences). Data analysis was performed using FlowJo software (Tree Star).
AZD5991 and navitoclax were purchased from Active Biochem; venetoclax and Q-VD-OPh were from MedChemExpress; AZD4320, IACS-010759 were from Selleck Chemicals. CCCP (carbonyl cyanide m-chlorophenyl hydrazine), NAC (N-Acetyl-L-cysteine) were from Sigma.

Liu T., Lam V., Thieme E., Sun D., Wang X., Xu F., Wang L., Danilova O.V., Xia Z., Tyner J.W., Kurtz S.E, & Danilov A.V. (2021). Pharmacologic targeting of Mcl-1 induces mitochondrial dysfunction and apoptosis in B-cell lymphoma cells in a TP53- and BAX-dependent manner. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(17), 4910-4922.

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Cell Line Maintenance and Compound Acquisition

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The pancreatic cancer cell line MIA PaCa-2 was purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan). The lung cancer cell lines LU65 and NCI-H23 were obtained from the RIKEN Cell Bank (Ibaraki, Japan) and American Type Culture Collection (Manassas, VA, USA), respectively. MIA PaCa-2 cells were maintained in Dulbecco’s modified Eagle medium supplemented with 10% fetal bovine serum (FBS), 0.1 mM nonessential amino acids, and 1 mM sodium pyruvate. NCI-H23 and LU65 cells were maintained in Roswell Park Memorial Institute 1640 medium supplemented with 10% FBS. In all culture media, 100 U/mL penicillin and 100 μg/mL streptomycin were added to prevent bacterial contamination. All cell culture reagents were purchased from Gibco, Thermo Fisher Scientific (Waltham, MA, USA). The cell lines were incubated in a humidified incubator at 37 °C under 5% CO₂ and 95% air atmosphere. All cells were verified through short-tandem repeat (STR) profiling and tested negative for mycoplasma contamination.
Sotorasib, ARS-1620, Q-VD-OPh (QVD), cobimetinib, N-acetyl-l-cysteine (NAC), and encorafenib were purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Chiou L.W., Chan C.H., Jhuang Y.L., Yang C.Y, & Jeng Y.M. (2023). DNA replication stress and mitotic catastrophe mediate sotorasib addiction in KRASG12C-mutant cancer. Journal of Biomedical Science, 30, 50.

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