Atezolizumab

Manufactured by Selleck Chemicals

Sourced in Germany, United States

Atezolizumab is a monoclonal antibody that binds to the PD-L1 protein, which is expressed on the surface of certain cells. It is used in research applications to study the immune system and potential therapeutic targets.

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Lab products found in correlation

Rpmi 1640, by Merck Group (3 mentions) Human nsclc cell lines, by American Type Culture Collection (2 mentions) Rabbit anti mouse igg, by Merck Group (2 mentions) Phospho mapk44 42, by Cell Signaling Technology (2 mentions) Mapk44 42, by Cell Signaling Technology (2 mentions) Mhc 1, by Cell Signaling Technology (2 mentions) Pd l1, by Cell Signaling Technology (2 mentions) Goat anti rabbit igg, by Merck Group (2 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions) Gflg tetrapeptide, by Peptron (1 mentions) Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions) Anti calreticulin antibody, by Abcam (1 mentions) Anti cd8 antibody, by Abcam (1 mentions) P sting, by Cell Signaling Technology (1 mentions) Human serum albumin (hsa), by Solarbio (1 mentions) Human ifn β elisa kit, by Beyotime (1 mentions) Anti foxp3, by Abcam (1 mentions) Goat anti rabbit igg h l alexa fluor 488, by Abcam (1 mentions) Mito tracker green, by Beyotime (1 mentions) Annexin 5 fitc apoptosis kit, by BD (1 mentions)

14 protocols using atezolizumab

Atezolizumab-Chlorin e6 Conjugate Protocol

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Atezolizumab was purchased
from Selleckchem Co. Ltd. (Huston, TX). Methoxy poly(ethylene glycol)-succinimidylglutarate
(MePEG–NHS, molecular weight (M.W.): 5000 g/mol) was purchased
from SunBio Co. Ltd. (Seoul, S. Korea). GFLG tetrapeptide was purchased
from Peptron Co (Daejeon, S. Korea). The dialysis membranes with molecular-weight
cutoffs (MWCO) of 1000, 2000, and 8000 g/mol were purchased from Spectra/ProTM
Membranes (New Brunswick, NJ). Chlorin e6 (Ce6) was purchased from
Frontier Scientific Inc. (Logan, UT). Phosphotungstic acid, N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide
hydrochloride (EDAC), N-hydroxysuccinimide (NHS),
triethylamine, cathepsin B (from bovine spleen), and dimethylsulfoxide
(DMSO) were purchased from Sigma Chem. Co. (St. Louis, MO). Organic
solvents such as methanol were used as high-performance liquid chromatography
grade or extra-pure grade.

Jeong Y.I., Yoo S.Y., Heo J, & Kang D.H. (2019). Chlorin e6-Conjugated and PEGylated Immune Checkpoint Inhibitor Nanocomposites for Pulmonary Metastatic Colorectal Cancer. ACS Omega, 4(20), 18593-18599.

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Apoptosis and Immune Response Induction

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IR780, HSA, zinc acetate, and sodium sulfide were purchased from Solarbio (Beijing, China). Fetal bovine serum (FBS) and Dulbecco’s modified Eagle medium (DMEM) were purchased from GIBCO (Grand Island, NY, USA). Mito-Tracker Green was purchased from Beyotime (Shanghai, China; #C1048). Annexin V-FITC Apoptosis Kit (BD, San Jose, CA, USA; #556557) was from Becton, Dickinson and Company (Franklin Lakes, NJ, USA). Anti-DFNA5/GSDME-N-terminal (Abcam, Cambridge, UK; #AB215191), anti-calreticulin antibody (Abcam, #ab92516), anti-high-mobility group box 1 (HMGB1) antibody (Abcam, #ab79823), anti-calreticulin antibody (Abcam, ab92516), goat anti-rabbit IgG H&L (Alexa Fluor 488) (Abcam, #ab150081), anti-CD8 antibody (Abcam, #ab217344), anti-FOXP3 (Abcam, #ab20034) were from Abcam. Cleaved caspase-3 (CST, Danvers, MA, USA; #9664T), cGAS (CST, #31659S), STING (CST, #50494S), and p-STING (CST, #50907) were from Cell Signaling Technology (Danvers, MA, USA). Atezolizumab (Selleck, Munich, Germany; #A2004) were from Selleck. Mouse IFN-β ELISA Kit (Beyotime, #PI568) and human IFN-β ELISA Kit (Beyotime, #PI572) were from Beyotime. SOSG9 (Meilunbio, Dalian, China; #MA0326) was from Meilunbio.

Yang J., Guo W., Huang R., Bian J., Zhang S., Wei T., He C., Hu Z., Li J., Zhou C, & Lu M. (2023). Self-assembled albumin nanoparticles induce pyroptosis for photodynamic/photothermal/immuno synergistic therapies in triple-negative breast cancer. Frontiers in Immunology, 14, 1173487.

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Cell Culture Conditions and Reagents

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HEK-293 T cells were maintained in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, C11995500BT). H460 (human large cell lung cancer), MKN-28 (human gastric carcinoma), SMMC-7721 (human hepatoma carcinoma), HeLa (human cervical cancer) and NALM6 (CD19⁺ acute lymphoblastic leukemia) cell lines were obtained from ATCC and maintained in RPMI-1640 (Gibco, C11975500BT). GL-expressing cell lines were generated through transduction of the parental cell line with a lentiviral supernatant containing GL and were sorted for GFP expression on a FACS Aria TM cell sorter (BD Biosciences). DMEM and RPMI-1640 medium were supplemented with 10% heat-inactivated FBS (Vigonob, XC6936T) and 1% penicillin/streptomycin. All cells were cultured at 37 °C in an atmosphere of 5% carbon dioxide. Atezolizumab (AZ) is a humanized anti-PD-L1 monoclonal antibody (Selleck).

Qin L., Zhao R., Chen D., Wei X., Wu Q., Long Y., Jiang Z., Li Y., Wu H., Zhang X., Wu Y., Cui S., Wei W., Yao H., Liu Z., Cao S., Yao Y., Zhang Z, & Li P. (2020). Chimeric antigen receptor T cells targeting PD-L1 suppress tumor growth. Biomarker Research, 8, 19.

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Metabolic Modulation of NK Cells

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Metabolic inhibitors were used on NK cells to study their effect on optical metabolic imaging measurements. 2DG (10 mM) (Sigma-Aldrich, D6134), etomoxir (10 μM) (Sigma-Aldrich, E1905), and oligomycin (1 μM) (Sigma-Aldrich, 75351) were used to perturb their respective pathways. To measure how immune checkpoint and IDO-1 inhibition affected NK cytotoxicity in the microdevice, the hydrogels were treated with 40 nM atezolizumab (Selleckchem, A2004) and 50 nM epacadostat (Selleckchem, S7910) for 7 days. After these treatments, cell viability was assessed as described above.

Ayuso J.M., Rehman S., Virumbrales-Munoz M., McMinn P.H., Geiger P., Fitzgerald C., Heaster T., Skala M.C, & Beebe D.J. (2021). Microfluidic tumor-on-a-chip model to evaluate the role of tumor environmental stress on NK cell exhaustion. Science Advances, 7(8), eabc2331.

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Combination Therapy Evaluation

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Cisplatin, Nivolumab, Atezolizumab, and Ipilimumab were purchased from Selleckchem (Houston, TX, USA). Solitomab was purchased from ProSci Inc. (Poway, CA, USA).

Xue B., Schüler J., Harrod C.M., Lashuk K., Bomya Z, & Hribar K.C. (2023). A Novel Hydrogel-Based 3D In Vitro Tumor Panel of 30 PDX Models Incorporates Tumor, Stromal and Immune Cell Compartments of the TME for the Screening of Oncology and Immuno-Therapies. Cells, 12(8), 1145.

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Culturing and Treating Primary Immune Cells

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Human PBMCs were cultured in Roswell Park Memorial Institute (RPMI) medium supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Biowest), 100 U penicillin–streptomycin (Gibco), and 2 mM L-Glutamine (Gibco). PDECs were cultured in MammoCult (STEMCELL Technologies), and the MammoCult media was supplemented with MammoCult proliferation supplement #05622 (STEMCELL Technologies), 20 µg/mL gentamicin (Sigma), 0.1 µg/mL amphotericin B (Biowest) and 10,000 U/mL penicillin/streptomycin (Lonza). Cells and PDECs were grown in a humidified incubator at 37°C under 5% CO₂, and atmospheric oxygen levels.
PDECs were treated with 25 ul/mL anti-CD3/CD28/CD2 (STEMCELL Technologies), 100 ug/mL atezolizumab (Selleck Chemicals), 50 ug/mL pembrolizumab (MedChem), 10–100 nM Venetoclax (MedChem Express), 5–10 mM metformin (MedChem Express), 10–50 nM paclitaxel (MedChem Express), 1–2.5 nM IACS-010759 (Selleck Chemicals), 10–30 nM rotenone (Sigma-Aldrich), and 10 nM-1 uM A-769662 (Sigma-Aldrich), 100 ng/mL lipopolysaccharide (LPS).

Turpin R., Liu R., Munne P.M., Peura A., Rannikko J.H., Philips G., Boeckx B., Salmelin N., Hurskainen E., Suleymanova I., Aung J., Vuorinen E.M., Lehtinen L., Mutka M., Kovanen P.E., Niinikoski L., Meretoja T.J., Mattson J., Mustjoki S., Saavalainen P., Goga A., Lambrechts D., Pouwels J., Hollmén M, & Klefström J. (2024). Respiratory complex I regulates dendritic cell maturation in explant model of human tumor immune microenvironment. Journal for Immunotherapy of Cancer, 12(4), e008053.

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Cytotoxicity Assay for T-cell-Mediated Tumor Killing

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T-cell-mediated tumor cell killing assay was performed as described previously.^{21 (link)} Tumor cells were pre-treated with MMC (0.4 μM) for 24 h. PBMCs were plated at a density of 1 × 10⁶/well in 6-well plates and then co-cultured with tumors cells at 4:1 ratio for 24 h. Anti-human PD-L1 antibody, atezolizumab (Selleck Chemicals, USA) (50 μg/ml) was added afterwards. The co-culture system was maintained for 5 days and the wells were washed with PBS twice to remove PBMCs. The adherent and surviving cancer cells were fixed and stained with crystal violet solution. Cancer cell viability was then estimated by colorimetric determination.

Luo M., Wang F., Zhang H., To K.K., Wu S., Chen Z., Liang S, & Fu L. (2020). Mitomycin C enhanced the efficacy of PD-L1 blockade in non-small cell lung cancer. Signal Transduction and Targeted Therapy, 5, 141.

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Isolation and Culture of NSCLC Cell Lines

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Human NSCLC cell lines were obtained by the American Type Culture Collection (ATCC, Manassas, Virginia, USA) and were cultured in RPMI-1640 (Sigma-Aldrich) medium which was supplemented with 10% foetal bovine serum (FBS; Life Technologies, Gaithersburg, Maryland, USA) in a humidified atmosphere with 5% CO₂. The identity of cell lines was confirmed by Short tandem repeat profiling (Promega). Cetuximab, atezolizumab and panitumumab were purchased from Selleck Chemicals (Munich, Germany). Avelumab, was provided by Merck (Darmstadt, Germany), as part of a Cooperative Research and Development Agreement. Peripheral blood mononuclear cells (PBMCs) from healthy donors (HD) or from NSCLC patients were isolated by Ficoll-Paque Plus (GE Healthcare). NK cells were obtained through magnetic separation by using the NK CELL isolation kit, HUMAN (Miltenyi Biotech 130-092-657) according to the manufacturer’s protocol.

Fasano M., Della Corte C.M., Di Liello R., Barra G., Sparano F., Viscardi G., Iacovino M.L., Paragliola F., Famiglietti V., Ciaramella V., Cimmino F., Capasso M., Iolascon A., Sforza V., Morabito A., Maiello E., Ciardiello F, & Morgillo F. (2020). Induction of natural killer antibody-dependent cell cytotoxicity and of clinical activity of cetuximab plus avelumab in non-small cell lung cancer. ESMO Open, 5(5), e000753.

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Immunological Profiling of NSCLC Cell Lines

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The human NSCLC cell lines were provided by American Type Culture Collection (ATCC, Manassas, VA, USA) and maintained in RPMI-1640 (Sigma-Aldrich) medium supplemented with 10% fetal bovine serum (FBS; Life Technologies, Gaithersburg, MD) in a humidified atmosphere with 5% CO2. The identity of all cell lines was confirmed by STR profiling (Promega) on an ad hoc basis prior to perform experiments.
Selumetinib (MEK-I, AZD6244) and atezolizumab were purchased from Selleck Chemicals, Munich, Germany. Avelumab, was provided by EMD Serono as a part of a Cooperative Research and Development agreement with our institution.
Primary antibodies for western blot analysis against phospho-MEK, MEK, phospho-MAPK44/42, MAPK44/42, PD-L1, phospho-STAT3 and MHC-I were obtained from Cell Signaling Technology; the following secondary antibodies from Bio-Rad were used: goat anti-rabbit IgG, rabbit anti-mouse IgG and monoclonal anti-β actin antibody from Sigma Chemical Co.

Della Corte C.M., Barra G., Ciaramella V., Di Liello R., Vicidomini G., Zappavigna S., Luce A., Abate M., Fiorelli A., Caraglia M., Santini M., Martinelli E., Troiani T., Ciardiello F, & Morgillo F. (2019). Antitumor activity of dual blockade of PD-L1 and MEK in NSCLC patients derived three-dimensional spheroid cultures. Journal of Experimental & Clinical Cancer Research : CR, 38, 253.

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Evaluating Atezolizumab's Impact on Osteosarcoma

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Human OS cell lines HOS and 143B were obtained from ATCC (Manassas, VA, USA). All cells were cultured in high glucose Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin at 37 °C. Different concentrations of atezolizumab (A2004, Selleck, USA) were added to the medium. SP600125, a JNK inhibitor (S1460, Selleck, USA) was added to the medium at 10 μM. Mito-TEMPO (SML0737, Sigma, USA) was added to the medium at 10 μM as a mitochondria-targeted antioxidant. To inhibit the autophagy induced by atezolizumab, OS cells were pre-treated with chloroquine (CQ) (C6628, Sigma, USA) for 16 h at 10 μM, and then atezolizumab and CQ were added together for another 24 h. We chose the OS cells treated with an apoptotic inducer (C0005, Beyotime, China) at 1:1000 for 24 h as the positive control of apoptosis-related proteins staining. And the positive control for autophagy was by starving the cells for 12 h.

Liu Z., Wang H., Hu C., Wu C., Wang J., Hu F., Fu Y., Wen J, & Zhang W. (2021). Targeting autophagy enhances atezolizumab-induced mitochondria-related apoptosis in osteosarcoma. Cell Death & Disease, 12(2), 164.

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