Carfilzomib

Manufactured by Apexbio

Sourced in United States

Carfilzomib is a proteasome inhibitor, a type of laboratory equipment used for protein analysis and drug discovery research. It functions by selectively and irreversibly binding to and inhibiting the proteasome, a large protein complex involved in the degradation of unwanted or damaged proteins within cells. Carfilzomib is commonly used in scientific research to study the role of the proteasome in cellular processes and to investigate the effects of proteasome inhibition on various biological systems.

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Lab products found in correlation

Bortezomib, by Selleck Chemicals (4 mentions) Bortezomib, by Apexbio (4 mentions) Hrp conjugated secondary antibody, by Jackson ImmunoResearch (2 mentions) Ethidium homodimer 1, by Thermo Fisher Scientific (2 mentions) Ixazomib, by Apexbio (2 mentions) Epoxomicin, by Apexbio (2 mentions) Antibody against baxα n20, by Santa Cruz Biotechnology (2 mentions) Ammonium chloride nh4cl, by Merck Group (1 mentions) Dithiothreitol (dtt), by Thermo Fisher Scientific (1 mentions) Thapsigargin, by Bio-Techne (1 mentions) Tunicamycin, by Bio-Techne (1 mentions) Mln4924, by LifeSensors (1 mentions) Bay11 7082, by Merck Group (1 mentions) Anti mouse and anti rabbit horseradish peroxidase hrp conjugated secondary antibodies, by Santa Cruz Biotechnology (1 mentions) Bortezomib, by Chemietek (1 mentions) Sc 514, by Santa Cruz Biotechnology (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Trolox, by Merck Group (1 mentions) Chloroquine, by Merck Group (1 mentions)

11 protocols using carfilzomib

Purification of Cellular Organelles

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DSP and ammonium chloride (NH₄Cl) were purchased from Sigma-Aldrich (St Louis, MO). BFA was purchased from BioLegend (San Diego, CA). Bortezomib was from Selleck Chemicals (Houston, TX). Carfilzomib was purchased from ApexBio Technology (Houston, TX).

Yamashita Y., Anczurowski M., Nakatsugawa M., Tanaka M., Kagoya Y., Sinha A., Chamoto K., Ochi T., Guo T., Saso K., Butler M.O., Minden M.D., Kislinger T, & Hirano N. (2017). HLA-DP84Gly constitutively presents endogenous peptides generated by the class I antigen processing pathway. Nature Communications, 8, 15244.

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Mycolactone Purification and Storage

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Mycolactone was purified from M. ulcerans bacterial pellets (strain 1615, ATCC 35840), then quantified by spectrophotometry, and stored in ethanol at −20°C protected from light. For in vivo experiments, a 4 mM stock was diluted in a NaCl solution (0.9% w/v) immediately before injection in animals. For in vitro experiments, a 1,000× working solution was prepared by dilution of the ethanol stock in DMSO and stored at −20°C, then thawed and diluted in culture medium immediately before use. BZ purchased from Alfa Aesar (#J60378) was resuspended in DMSO to yield a 10 mM solution stored at −20°C. A 10 mM solution of carfilzomib in DMSO was purchased from ApexBio (#A1933) and stored at −20°C. BZ and carfilzomib were thawed and diluted in culture medium immediately before use. Thapsigargin (Tocris, #1138) and tunicamycin (Tocris, #3516) were resuspended in DMSO to yield a 10 mM solution stored at −20°C; and dithiothreitol (DTT, 1 M solution) was purchased from Invitrogen (#P2325) and stored at −20°C.

Domenger A., Choisy C., Baron L., Mayau V., Perthame E., Deriano L., Arnulf B., Bories J., Dadaglio G, & Demangel C. (2022). The Sec61 translocon is a therapeutic vulnerability in multiple myeloma. EMBO Molecular Medicine, 14(3), e14740.

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Apoptosis Regulation by Proteasome Inhibitors

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Bortezomib, carfilzomib, epoxomicin, and ixazomib were obtained from ApexBio. Inhibitors against caspase 3 (Z-DEVD-FMK), caspase 6 (Z-VEID-FMK), caspase 8 (z-VETD-FMK), caspase 9 (z-LEHD-FMK), and caspase 12 (ATAD-FMK) were obtained from Calbiochem. Ethidium-homodimer-1 was obtained from Invitrogen. Antibody against BaxΔ2 was generated previously [20 ]. Antibodies against actin, cleaved caspase 8, LC3B, and Bcl-2 were obtained from Cell Signaling. Antibody against Baxα (N20) was from Santa Cruz Biotechnology, and HRP-conjugated secondary antibody was from Jackson.

Mañas A., Chen W., Nelson A., Yao Q, & Xiang J. (2017). BaxΔ2 sensitizes colorectal cancer cells to proteasome inhibitor-induced cell death. Biochemical and biophysical research communications, 496(1), 18-24.

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Reversible Proteasomal Inhibition in Cardiomyocytes

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MG132, Bortezomib/Velcade, NAE inhibitor MLN 4924, AAA-ATPase p97 inhibitor DBeQ were purchased from LifeSensors (Malvern, PA). Carfilzomib were obtained from APExBIO technology. All inhibitors were dissolved in DMSO. The inhibitors were added to the muscle cells after 4 days of culture. To reverse the drug effects, the treated cells were washed, trypsinized, spun down, rinsed with control medium, and then replated onto gelatin-coated dishes (hiPSC) or collagen-coated dishes (chick) in control medium. The control and different previously UPS inhibitor treated cardiomyocytes were allowed spread in culture over two days for recovery, and then processed for immunostainings to detect reversibility.

Wang J., Fan Y., Wang C., Dube S., Poiesz B.J., Dube D.K., Ma Z., Sanger J.M, & Sanger J.W. (2022). Inhibitors of the Ubiquitin Proteasome System block myofibril assembly in cardiomyocytes derived from chick embryos and human pluripotent stem cells. Cytoskeleton (Hoboken, N.J.), 78(10-12), 461-491.

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Apoptosis Regulation by Proteasome Inhibitors

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Proteomic Analysis of NF-κB Pathway

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Antibodies against human CXCR1 (sc-7303), CXCR2 (sc-7304), IKKα (sc-7218), IKKβ (sc-8014), IKKε (sc-376114), p65 NFκB (sc-372), IκBα (sc-371), and histone H3 (sc-8654) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibody against lactate dehydrogenase (LDH; 20-LG22) was from Fitzgerald Industries International (North Acton, MA, USA), and actin antibody was from Sigma (St Louis, MO, USA). Horseradish peroxidase (HRP)-conjugated anti-rabbit and anti-mouse secondary antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA).
Bortezomib was from ChemieTek (Indianapolis, IN, USA), and carfilzomib was from ApexBio (Houston, TX, USA). Bay-117082 was purchased from Sigma, and SC514 was from Santa Cruz Biotechnology. All other reagents were molecular biology grade and were from Sigma (St Louis, MO).

Uddin M.M., Zou Y., Sharma T., Gatla H.R, & Vancurova I. (2018). Proteasome inhibition induces IKK-dependent interleukin-8 expression in triple negative breast cancer cells: Opportunity for combination therapy. PLoS ONE, 13(8), e0201858.

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Assessing Proteasome Inhibitor Sensitivity

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Small interference double‐strand RNA (siRNA pool, M‐006022‐01‐0005) against the PSMB8 gene and nontarget siRNA as control were purchased from Dharmacon. The cell viability assay was performed by seeding 1,000 cells in 96‐well plates with complete media and detected at 0, 24, 48, 72 and 128 hr. After adding 3‐(4,5‐dimethylthiazol‐2‐yl)‐2‐(4‐sulfophenyl)‐2 H‐tetrazolium reagents (Promega) for 1 hr, each well was measured the absorbance using OD 490 nm in MultiSKAN Ex ELISA reader (Thermo Scientific Pierce). The chemosensitivity assay was performed by seeding 1,000 cells in 96‐well plates with complete media and detected in adding 0, 1, 2, 4, 8, 16, 32, 64, 128 and 256 nM Carfilzomib or Bortezomib (Apexbio). The Carfilzomib or Bortezomib are proteasome inhibitors. Carfilzomib targets PSMB 1, 2, 5, 8, 9 and 10. Bortezomib specifically inhibits PSMB1 and PSMB5.

Liew P., Huang R., Weng Y., Fang C., Hui‐Ming Huang T, & Lai H. (2018). Distinct methylation profile of mucinous ovarian carcinoma reveals susceptibility to proteasome inhibitors. International Journal of Cancer, 143(2), 355-367.

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Multimodal Combination Therapy Screening

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Chloroquine (20 μM), Buthionine sulfoximine (BSO, 50 μM), Arsenic trioxide (As₂O₃,1 μM), N-Acetyl-L-cysteine (NAC, 2 mM), Palbociclib (1 μM) and 4’,6-diamidino-2-phenylindole (DAPI, 1 μg/ml) were from Sigma-Aldrich. Lapatinib (1 μM), Bortezomib (50 nM), PF4708671 (10 μM), LY2584702 (1 μM), and MK-2206 (1 μM) were from Selleckchem. Trolox (50 μM, EMD Millipore). MG132 (1 μM, Tocris). Carfilzomib (50 nM, APExBIO). Selonsertib (10 μM) and staurosporine (10 μM) were from Adooq. Doxycycline (1 μg/ml) was from Research Products International. Hoechst 33342 (5 μg/ml) was from ThermoFisher Scientific.

Kim L.M., Kim P.Y., Gebreyohannes Y.K, & Leung C.T. (2022). Sustained oncogenic signaling in the cytostatic state enables targeting of non-proliferating persistent cancer cells. Cancer research, 82(17), 3045-3057.

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Propagation of Ehrlichia chaffeensis in THP-1 Cells

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Ehrlichia chaffeensis (Arkansas strain) was propagated in a human monocytic cell line (THP-1; ATCC, Manassas, VA). THP-1 cells were maintained in RPMI 1640 with HEPES (25 mM) (Invitrogen; Carlsbad, CA) supplemented with 10% fetal bovine serum (FBS)(HyClone, Logan, UT), 5 mM L-glutamine, 1% sodium pyruvate, and 12.5 ml 10% glucose (Sigma; St. Louis, MO) at 37°C in a 5% CO₂ atmosphere. E. chaffeensis infection was maintained by subculturing infected cells with uninfected. HeLa cells (human cervical epithelial; ATCC) for transfection were grown in MEM (Invitrogen) supplemented with 10% FBS (HyClone) and maintained in a 5% CO₂ atmosphere. When stated cells were incubated with 10 mM bortezomib (ApexBio; Houston, TX), PYR41 (Thermo Fisher; Boston, MA), Heclin (Sigma), carfilzomib (ApexBio) or vehicle (DMSO) for 12–24 h before harvesting cells for lysate or luciferase expression measurement.

Farris T.R., Zhu B., Wang J.Y, & McBride J.W. (2018). Ehrlichia chaffeensis TRP32 Nucleomodulin Function and Localization Is Regulated by NEDD4L-Mediated Ubiquitination. Frontiers in Cellular and Infection Microbiology, 7, 534.

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In Vitro Characterization of Human Lung Cancer Cell Lines

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Human lung cancer cell lines (NCI‐H820, NCI‐H1975) were purchased from the American Type Culture Collection (ATCC). All cell lines were cultured as previously described [18], and mycoplasma testing was routinely performed using e‐Myco™ plus Mycoplasma PCR Detection Kit (Intron, Korea), verifying that the cells were mycoplasma free. Anti‐PARP, anticleaved caspase 3, anti‐AKT, anti‐phospho‐AKT, anti‐ERK, and anti‐phospho‐ERK antibodies were purchased from Cell Signaling Technology. Erlotinib, SJ‐172550, nutlin‐3, and carfilzomib were obtained from APExBIO. Bortezomib, quinacrine, brefeldin A, and ouabain were purchased from Selleckchem (Houston, TX, USA), and N‐ethylmaleimide was obtained from the Sigma‐Aldrich Corporation (Sigma, St. Louis, MO, USA).

Lee J., Choi A., Cho S., Jun Y., Na D., Lee A., Jang G., Kwon J.Y., Kim J., Lee S, & Lee C. (2020). Genome‐scale CRISPR screening identifies cell cycle and protein ubiquitination processes as druggable targets for erlotinib‐resistant lung cancer. Molecular Oncology, 15(2), 487-502.

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