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Navitoclax

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Navitoclax is a small molecule designed for laboratory use. It functions as a selective inhibitor of the Bcl-2 family of anti-apoptotic proteins.

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Lab products found in correlation

Phosal 50 pg, by Lipoid (2 mentions) Bcl xl, by Cell Signaling Technology (2 mentions) Mcl 1, by Cell Signaling Technology (2 mentions) Peg400, by Merck Group (2 mentions) Panobinostat, by Selleck Chemicals (1 mentions) Panobinostat, by Merck Group (1 mentions) Quisinostat, by Selleck Chemicals (1 mentions) Mannitol, by Merck Group (1 mentions) Navitoclax, by Thermo Fisher Scientific (1 mentions) A 1331852, by Chemietek (1 mentions) Napabucasin, by Selleck Chemicals (1 mentions) Napabucasin, by Merck Group (1 mentions) Peg300, by Merck Group (1 mentions) Hydroxypropyl β cyclodextrin, by Merck Group (1 mentions) Tween 80, by Merck Group (1 mentions) Venetoclax, by Chemietek (1 mentions) Digitonin, by Merck Group (1 mentions) Bcl w, by Cell Signaling Technology (1 mentions) Obatoclax, by Selleck Chemicals (1 mentions) Vdac2, by Cell Signaling Technology (1 mentions)

4 protocols using navitoclax

1

Drug Formulation and Preparation Protocol

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Napabucasin, panobinostat, and quisinostat were purchased from Selleckchem and A1331852 and Navitoclax from Chemietek, diluted in DMSO to make stock solutions, aliquoted and stored at −80C. Napabucasin was formulated by heating to 50C for 10 minutes and then sequentially adding 45% PEG300 (Sigma), 5% Tween80 (Sigma) and 45% sterile water, with vortexing after adding each component (Selleckchem). panobinostat was formulated by sequentially adding 48% PEG300 (Sigma), 2% Tween80 (Sigma) and 48% sterile water, with vortexing after adding each component (Selleckchem). quisinostat was formulated in 10% hydroxypropyl-b-cyclodextrin (Sigma), 25 mg/ml mannitol (Sigma), in sterile water(45 (link)). A1331852 was formulated by sequentially adding 10% Ethanol (Fisher), 60% Phosal 50 PG (Lipoid), and 30% PEG400 (Sigma), and vortexing (MedChemExpress). Navitoclax was formulated in 10% ethanol (Fisher), 30% PEG400 (Sigma), and 60% Phosal 50 PG (Lipoid), with vortexing after adding each component (MedChemExpress). Sorafenib was formulated in 90% corn oil (Selleckchem) with vortexing (MedChemExpress). Working solutions were made fresh prior to administration.
Lalazar G., Requena D., Ramos-Espiritu L., Ng D., Bhola P.D., de Jong Y.P., Wang R., Narayan N.J., Shebl B., Levin S., Michalidis E., Kabbani M., Vercauteren K.O., Hurley A.M., Farber B.A., Hammond W.J., Saltsman JA I.I.I., Weinberg E.M., Glickman J.F., Lyons B.A., Ellison J., Schadde E., Hertl M., Leiting J.L., Truty M.J., Smoot R.L., Tierney F., Kato T., Wendel H.G., LaQuaglia M.P., Rice C.M., Letai A., Coffino P., Torbenson M.S., Ortiz M.V, & Simon S.M. (2021). Identification of Novel Therapeutic Targets for Fibrolamellar Carcinoma Using Patient Derived Xenografts and Direct from Patient Screening. Cancer discovery, 11(10), 2544-2563.
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2

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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3

Formulation and Dilution of Anticancer Agents

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Irinotecan (Selleckchem, S2217) and navitoclax (ChemieTek, CT-A263) were diluted in DMSO to make stock solutions, were aliquoted, and were stored at –80°C. DT2216, both the i.p. (nonclinical) and i.v. (clinical) formulations, were formulated as previously reported (20 (link)). The clinical formulation of DT2216 was diluted 1:1 in 5% dextrose to achieve a higher volume of injection in mice. Irinotecan was formulated in 99.5% of 0.9% sodium chloride (Thermo Fisher Scientific) and vortexed. navitoclax was formulated in 10% ethanol (Thermo Fisher Scientific), 30% PEG-400 (MilliporeSigma), and 60% Phosal 50PG (Lipoid), with vortexing after adding each component. Working solutions of Irinotecan and navitoclax were made fresh prior to each administration.
Shebl B., Ng D., Lalazar G., Rosemore C., Finkelstein T.M., Migler R.D., Zheng G., Zhang P., Jiang C.S., Qureshi A., Vaughan R., Yarchoan M., de Jong Y.P., Rice C.M., Coffino P., Ortiz M.V., Zhou D, & Simon S.M. (2022). Targeting BCL-XL in fibrolamellar hepatocellular carcinoma. JCI Insight, 7(17), e161820.
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4

Apoptosis Signaling Pathway Assay Protocol

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Reagents were obtained from the following suppliers: lipids and extruder from Avanti Polar Lipids, CM5 biosensor chips from GE Healthcare, Polysorbate 20 from Biacore AB, Q-VD-OPh from SM Biochemicals (Anaheim, CA), glutathione from Sigma, Ni2+-NTA-agarose from Novagen, fluorescein isothiocyanate (FITC)-labeled dextran 10 from Invitrogen, navitoclax, and S63845 from Chemietek (Indianapolis, IN). Antibodies to the following antigens were purchased from the indicated suppliers and used with the indicated dilutions: Hsp60 (cat. #4870 S, 1:1000), EGFP (#2555 S, 1:1000), BAX (#2772 S, 1:1000), BIM (#2933 S, 1:1000), BID (#2002S, 1:1000), BCLXL(#2764 S, 1:1000), and MCL1 (#4572 S, 1:1000) from Cell Signaling Technology; cytochrome c (#556433, 1:1000) from BD Biosciences; BAK N-term antibody (#06-536, 1:1000) and BAK Ab-1 antibody (TC-100) from Millipore; BCL2 antibody from DAKO (#M0887, 1:1000); NOXA antibody from Enzo Life Sciences (#ALX-804-408-c100, 1:1000); BMF antibody from Proteintech (#18298-1-AP, 1:1000) and actin (goat polyclonal, I-19, #sc-1615, 1:500) and PUMA (#sc-374223, 1:1000) from Santa Cruz Biotechnology. Anti-S peptide antibody was raised in our laboratory and has been validated42 (link). BH3 peptides were generated by solid phase synthesis at GL Biochem Ltd. (Shanghai, China).
Ye K., Meng W.X., Sun H., Wu B., Chen M., Pang Y.P., Gao J., Wang H., Wang J., Kaufmann S.H, & Dai H. (2020). Characterization of an alternative BAK-binding site for BH3 peptides. Nature Communications, 11, 3301.
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