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Venetoclax

Manufactured by Chemietek
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Venetoclax is a small molecule that binds to the BCL-2 protein, which is involved in regulating cell death. It is used in the laboratory setting for research purposes.

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Lab products found in correlation

Verapamil, by Merck Group (2 mentions) Dasatinib, by LC Laboratories (2 mentions) Dimethyl sulfoxide (dmso), by Merck Group (2 mentions) Amg 176, by Chemietek (2 mentions) Vincristine, by Selleck Chemicals (2 mentions) Bcl xl, by Cell Signaling Technology (2 mentions) Mcl 1, by Cell Signaling Technology (2 mentions) Triton x 100, by Merck Group (1 mentions) 3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt dimethyl sulfoxide dmso, by Merck Group (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Trypsin, by Corning (1 mentions) Paclitaxel, by Merck Group (1 mentions) Cisplatin, by Merck Group (1 mentions) Ko143, by Merck Group (1 mentions) Mitoxantrone, by Merck Group (1 mentions) Topotecan, by Merck Group (1 mentions) Alexa fluor 488 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Sn 38, by Merck Group (1 mentions) Quizartinib, by LC Laboratories (1 mentions) Entospletinib, by Selleck Chemicals (1 mentions)

Close spelling variants of this product

Venetoclax (ABT-199) (2 citations)

10 protocols using venetoclax

1

Assessing Anticancer Drug Interactions

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Anticancer drugs and positive reversal agents: Venetoclax was purchased from ChemieTek (Indianapolis, IN, USA). Paclitaxel, mitoxantrone, topotecan, SN-38, verapamil, KO143, and cisplatin were purchased from Sigma Co (St. Louis, MO, USA). Cell culture: Bovine serum (BS), fetal bovine serum (FBS), Dulbecco’s modified eagle’s medium (DMEM) and trypsin (0.25%) were purchased from Corning Inc. (Corning, NY, USA). Cell viability assay: dimethylsulfoxide (DMSO), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma Co. Immunoblotting: Human monoclonal antibodies for ABCG2 and GAPDH were purchased from Millipore (Billerica, MA, USA). Alexa Fluor 488 conjugated secondary antibody was purchased from Thermo Fisher Scientific Inc (Rockford, IL, USA). Triton X-100, 4′,6-diamidino-2-phenylindole (DAPI) were purchased from Sigma Co. Other chemicals: [3H]-labeled mitoxantrone (2.5 Ci/mmol) was purchased from Moravek Biochemicals (Brea, CA, USA). All other chemicals were purchased from Sigma Co.
Wang J.Q., Li J.Y., Teng Q.X., Lei Z.N., Ji N., Cui Q., Zeng L., Pan Y., Yang D.H, & Chen Z.S. (2020). Venetoclax, a BCL-2 Inhibitor, Enhances the Efficacy of Chemotherapeutic Agents in Wild-Type ABCG2-Overexpression-Mediated MDR Cancer Cells. Cancers, 12(2), 466.
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2

Inhibitor Compound Preparation Protocol

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ARQ 531 was synthesized and provided by ArQule, Inc., (Burlington, MA). Quizartinib, midostaurin, and dasatinib were purchased from LC Laboratories (Woburn, MA). Gilteritinib and venetoclax were purchased from Chemietek (Indianapolis, IN). Entospletinib (GS-9973) was purchased from Selleckchem (Houston, TX). All chemical compounds were dissolved in Dimethyl sulfoxide (DMSO) to obtain a 10 mM stock solution.
Elgamal O.A., Mehmood A., Jeon J.Y., Carmichael B., Lehman A., Orwick S.J., Truxall J., Goettl V.M., Wasmuth R., Tran M., Mitchell S., Lapalombella R., Eathiraj S., Schwartz B., Stegmaier K., Baker S.D., Hertlein E, & Byrd J.C. (2020). Preclinical efficacy for a novel tyrosine kinase inhibitor, ArQule 531 against acute myeloid leukemia. Journal of Hematology & Oncology, 13, 8.
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3

Preparation of Compound Solutions

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Venetoclax (>99.9%, Chemietek, catalog no. CT-A199), A-1155463 (99.5%, Chemietek, catalog no. CT-A115) and NVP-CGM097 (100% optically pure, Chemietek, catalog no. CT-CGM097) were used directly without further purification. Venetoclax, A-1155463 and NVP-CGM097 were each dissolved in DMSO as 10 mM stocks. Stocks were aliquoted and stored at −20 °C until use.
Shui S., Scheller L, & Correia B.E. (2023). Protein-based bandpass filters for controlling cellular signaling with chemical inputs. Nature Chemical Biology, 20(5), 586-593.
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4

Synergistic Evaluation of AMG-176 and Venetoclax

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AMG-176 and venetoclax were purchased from Chemietek (Indianapolis, IN) and Xcessbio Company (San Diego, CA), respectively. For both drugs, stock solutions were made in dimethyl sulfoxide (DMSO; Sigma-Aldrich). The pan caspase inhibitor, quinoline-Val-Asp (Ome)-CH2-O-Ph (Q-VD-Oph) was obtained from APExBIO (Houston, TX). Typically, incubations were performed for 24 hours and for all treatments, time-matched, DMSO-treated (i.e., vehicle only) cells were used as controls. MEC1 cell line was purchased from Deutsche Sammlung von Mikroorganismen und Zellkulturen (DMSZ, Braunschweig, Germany). It was authenticated using short-tandem repeat profiling and regularly tested for Mycoplasma contamination using Mycoalert Mycoplasma detection kit (Lonza Group Ltd, Basel, Switzerland). We use cells between 5th and 20th passage. The cells were used as a reference protein concentration for the Bcl-2 family proteins.
Yi X., Sarkar A., Kismali G., Aslan B., Ayres M., Iles L.R., Keating M.J., Wierda W.G., Long J.P., Sabrina Bertilaccio M.T, & Gandhi V. (2020). AMG-176, an Mcl-1 antagonist, shows preclinical efficacy in chronic lymphocytic leukemia. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(14), 3856-3867.
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5

Investigating IP3 Signaling and Apoptosis

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Reagents were as follows: ethylene glycol tetraacetic acid (EGTA) (Acros Organics, Geel, Belgium, 409910250), Fura-2 AM (Biotium, Kampenhout, Belgium, 50033), Annexin V-Fluorescein isothiocyanate (FITC) (Becton Dickinson, Franklin Lakes, NJ, USA, 556419), 7-aminoactinomycin D (7-AAD) (Becton Dickinson, 555815), U73122 (Enzo Life Sciences, Farmingdale, NY, USA, BML-ST391-0005), U73343 (Enzo Life Sciences, BML-ST392-0005), venetoclax (ChemieTek, Indianapolis, IN, USA, CT-A199), anti-human IgG/M (Jackson ImmunoResearch, West Grove, PA, USA, 109-006-127). The following antibodies were used: anti-IP3R2 (Abiocode, Agoura Hills, CA, USA, R2872-3); anti-calnexin (Enzo Life Sciences, Farmingdale, NY, USA, ADI-SPA-865-D); anti-Bcl-2 (Santa Cruz Biotechnology, Dallas, TX, USA, sc7382HRP); anti-Bim (Bioké, Leiden, The Netherlands, 2819 S); anti-GAPDH (Sigma-Aldrich, St. Louis, MO, USA, G8795); anti-vinculin (Sigma-Aldrich, Munich, Germany, V9131). The sequences of the peptides used in this study were: BIRD-2 (RKKRRQRRRGGNVYTEIKCNSLLPLAAIVRV) and TAT-Ctrl (RKKRRQRRRGGSIELDDPRPR). These peptides were synthesized by LifeTein (South Plainfield, New Jersey, USA) with a purity of at least 85%. The IP3 sponge (pEF-GSTm49-IRES-GFP) is a protein constructed from the IP3-binding core of the type 1 IP3R with a single amino acid substitution (R441Q) that has a very high affinity for IP3 [29 (link)].
Bittremieux M., La Rovere R.M., Akl H., Martines C., Welkenhuyzen K., Dubron K., Baes M., Janssens A., Vandenberghe P., Laurenti L., Rietdorf K., Morciano G., Pinton P., Mikoshiba K., Bootman M.D., Efremov D.G., De Smedt H., Parys J.B, & Bultynck G. (2018). Constitutive IP3 signaling underlies the sensitivity of B-cell cancers to the Bcl-2/IP3 receptor disruptor BIRD-2. Cell Death and Differentiation, 26(3), 531-547.
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6

Diverse Compound Sourcing for Research

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Vincristine (VCR, #S2141), afatinib (#S1011), tariquidar (#S8028), lapatinib (#S2111), ceritinib (#S7083), copanlisib (#S2802), and decitabine (#S1200) were purchased from Selleckchem (Houston, TX, USA). Verapamil (#V4629), dactinomycin (#A4262), and hydroxychloroquine (#C6628) were purchased from Sigma–Aldrich (Merck KGaA, Darmstadt, Germany). BKM120 (#11587) and panobinostat (#13280) were purchased from Cayman Chemicals (Ann Arbor, MI, USA). Dasatinib (#D‐3307), everolimus (#E‐4040), and olaparib (#O‐9201) were purchased from LC Laboratories (Woburn, MA, USA). Staurosporine (STS, #HY‐15141) and larotrectinib (#HY‐12866) were purchased from Medchem Express (Monmouth Junction, NJ, USA). Bortezomib (#CT‐BZ001), trametinib (#CT‐GSK112), and venetoclax (#CT‐A199‐2) were purchased from ChemieTek (Indianapolis, IN, USA). The ERBB4 blocking antibody (#MA513016) was purchased from Invitrogen (Thermo Fisher Scientific Inc.).
Rösch L., Herter S., Najafi S., Ridinger J., Peterziel H., Cinatl J., Jones D.T., Michaelis M., Witt O, & Oehme I. (2022). ERBB and P‐glycoprotein inhibitors break resistance in relapsed neuroblastoma models through P‐glycoprotein. Molecular Oncology, 17(1), 37-58.
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7

Apoptosis Pathway Protein Analysis

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Reagents were obtained as follows: glutathione (GSH)-agarose from Thermo Scientific, Ni2+-NTA-agarose from Novagen, navitoclax and venetoclax from ChemieTek, A1210477 from Active Biochem, digitonin from Sigma-Aldrich, and obatoclax from Selleck Chemicals. BAK BH3 and BIM BH3 peptides were produced by solid-phase synthesis in the Mayo Clinic Proteomics Core. All other reagents were obtained as described (Dai et al. 2011 (link), 2014 (link)).
Antibodies were from the following suppliers: murine monoclonal anti-cytochrome c from BD Biosciences; murine monoclonal anti-BCL2 from Dako; goat anti-β-Actin and rabbit anti-PUMA antibodies from Santa Cruz Biotechnology; rabbit anti-BAK, mouse anti-BAK Ab-1, and rabbit anti-VDAC1 from Millipore; mouse anti-BCLB and rabbit monoclonal anti-BCL2A1 from Abcam; and rabbit antibodies to BAX, BCLXL, BIM, MCL1, BCLW, VDAC2, HSP60, GAPDH, and GFP from Cell Signaling Technology. Anti-S peptide antibody was raised as described (Hackbarth et al. 2004 (link)). Rat monoclonal anti-BID antibody was a kind gift from David Huang (Walter and Eliza Hall Institute, Melbourne, Australia).
Dai H., Ding H., Meng X.W., Peterson K.L., Schneider P.A., Karp J.E, & Kaufmann S.H. (2015). Constitutive BAK activation as a determinant of drug sensitivity in malignant lymphohematopoietic cells. Genes & Development, 29(20), 2140-2152.
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8

Apoptosis Pathway Protein Reagents

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Reagents were obtained as follows: Navitoclax and A1210477 from MedChemExpress; venetoclax and S63845 from Chemietek; topotecan from Toronto Research Chemicals; olaparib, etoposide, and vincristine from Selleck Chemicals; 5-fluorouracil from Tokyo Chemical Industry; and CHAPS from Millipore-Sigma. Antibodies were purchased from the following suppliers: anti-PUMA (#sc-374223, 1:1000) and anti-actin (goat polyclonal, I-19, #sc-1615, 1:500) antibodies from Santa Cruz Biotechnology; anti-NOXA antibody from ENZO Life Sciences (#ALX-804-408-c100, 1:1000); anti-BCL2 antibody from DAKO (#M0887, 1:1000); anti-BAK antibody from Millipore (#06-536, 1:1000) and antibodies to BAX (#2772S, 1:1000), BCLXL (#2764S, 1:1000), BIM (#2933S, 1:1000), MCL1 (#4572S, 1:1000), caspase-3 (#9662, 1:1000), caspase-9 (#9502, 1:1000), and tubulin (#2148, 1:1000) from Cell Signaling Technology. Rat anti-BID antibody was a kind gift from David Huang (Walter & Eliza Hall Institute, Melbourne, Australia).
Liu D., Hou X., Wu W., Zanfagnin V., Li Y., Correia C., Zhao Z., Zhao C., Liu Z., Zhang T., Fang Z., Wang H., Xu C., Weroha S.J., Kaufmann S.H, & Dai H. (2021). Constitutive BAK/MCL1 complexes predict paclitaxel and S63845 sensitivity of ovarian cancer. Cell Death & Disease, 12(8), 789.
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9

Combination Treatment of AMG-176 and Venetoclax

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AMG-176 and venetoclax were purchased from Chemietek (Indianapolis, IN) and Xcessbio Company (San Diego, CA), respectively. For both drugs, stock solutions were made in dimethyl sulfoxide (DMSO; Sigma-Aldrich). All incubations were for 24 h at the indicated drug concentration alone or in combination. DMSO only was used as a vehicle control.
Yi X., Jain N., Iles L.R., Ayres M.L., Wierda W.G, & Gandhi V. (2022). Targeting Mcl-1 by AMG-176 During Ibrutinib and Venetoclax Therapy in Chronic Lymphocytic Leukemia. Frontiers in Oncology, 12, 833714.
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10

Synthesis and Characterization of ISC-4 for Leukemia Research

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ISC-4 was synthesized using our previously described method at Penn State College of Medicine Organic Synthesis Core (16 (link)). Purity of ISC-4 (≥97%) was quantified by analytical high performance liquid chromatography (HPLC) analysis. Cytarabine (AraC, NDC 61703-305-38), Azacitidine (Aza, NDC 63323-771-39), and Daunorubicin (DNR, NDC 0703-5233-11) were purchased from the Penn State Hershey Medical Center Pharmacy. AraC was stored at room temperature, DNR at 4°C, and Azacitidine at −80°C. Venetoclax was purchased from ChemieTek (Indianapolis). ISC-4, Venetoclax and GDC-0941 were reconstituted in DMSO and aliqouts were stored at −20°C. GDC-0941 (catalog # S1065) was purchased from Selleck Chemicals and reconstituted in DMSO. Aliquots were stored in −80°C.
Annageldiyev C., Tan S.F., Thakur S., Dhanyamraju P.K., Ramisetti S.R., Bhadauria P., Schick J., Zeng Z., Sharma V., Dunton W., Dovat S., Desai D., Zheng H., Feith D.J., Loughran TP J.r., Amin S., Sharma A.K., Claxton D, & Sharma A. (2020). The PI3K/AKT Pathway Inhibitor ISC-4 Induces Apoptosis and Inhibits Growth of Leukemia in Preclinical Models of Acute Myeloid Leukemia. Frontiers in Oncology, 10, 393.
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