Ipilimumab

Manufactured by Selleck Chemicals

Sourced in United States

Ipilimumab is a laboratory reagent used for scientific research. It is a monoclonal antibody that targets the immune checkpoint protein CTLA-4. Ipilimumab is commonly used in studies related to cancer immunotherapy and the modulation of the immune system.

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6 protocols using ipilimumab

Evaluating Celecoxib and Ipilimumab Synergy

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We performed the drug assessment using selective COX-2 inhibitor celecoxib (SML3031, Merck, Darmstadt, Germany) at a concentration of 5 mM and the anti-CTLA-4 monoclonal antibody ipilimumab (A2001, Selleckchem, Houston, TX, USA) at a concentration of 80 nM. These dose concentrations of celecoxib and ipilimumab represent values usually achieved clinically.

Gómez-Valenzuela F., Wichmann I., Suárez F., Kato S., Ossandón E., Hermoso M., Fernández E.A, & Cuello M.A. (2023). Cyclooxygenase-2 Blockade Is Crucial to Restore Natural Killer Cell Activity before Anti-CTLA-4 Therapy against High-Grade Serous Ovarian Cancer. Cancers, 16(1), 80.

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Organoid Chemotherapy and Immunotherapy Evaluation

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Organoids were subsequently treated after 7 days of culture. Chemotherapies included cisplatin (1, 10, 100 µM) (232120, Sigma Aldrich), doxorubicin (0.1, 1, 10 µM) (S1208, Selleckchem), vincristine (0.1, 1, 10 µM) (V8879 Sigma Aldrich), doxorubicin with cisplatin (0.1/1, 1/10, 10/100 µM), cisplatin with etoposide (S1125, Selleckchem) (1/0.1, 10/1, 100/10 µM) (Selleckchem), and doxorubicin with vincristine (0.1/0.1, 1/1, 10/10 µM). For immunotherapy treatment, 100 nM of pembrolizumab (A2002, Selleckchem), ipilimumab (A2001, Selleckchem) or nivolumab (A2005, Selleckchem) was used on both iPTOs and PTOs in parallel treatments. Media was removed from the wells and drug solutions mixed in culture media were added. Organoids remained in treated media solution for 72 h prior to endpoint viability assessment.

Forsythe S.D., Erali R.A., Laney P., Sivakumar H., Li W., Skardal A., Soker S, & Votanopoulos K.I. (2022). Application of immune enhanced organoids in modeling personalized Merkel cell carcinoma research. Scientific Reports, 12, 13865.

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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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Sarcoma Organoid Drug Screening

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Therapy screening was initiated on day 7 following organoid biofabrication. Treatments were tailored for each sarcoma subtype based on preoperative biopsies or final surgical pathology. The number of therapies and doses involved varied based on available cells. The therapies used for sarcoma organoids were doxorubicin (0.1, 1, 10 μM) [S1208, Selleckchem, Houston, TX, USA], ifosfamide (2, 20, 200 μM) [I4909, Sigma-Aldrich], temozolomide (10, 100, 1000 μM) [T2577, Sigma-Aldrich], imatinib mesylate (1, 10, 100 μM) [STI571, Selleckchem], regorafenib (0.1, 1, 10 μM) [S1178, Selleckchem], gemcitabine (1, 10, 100 μM) [G6423, Sigma-Aldrich], olaparib (0.1, 1, 10 μM) [S1060, Selleckchem], 100 nM pembrolizumab (A2005, Selleckchem,), 100 nM nivolumab (A2002, Selleckchem), and 100 nM ipilimumab (A2001, Selleckchem). Drug containing-media was added to organoid wells for 72 h, followed by viability assays.

Forsythe S.D., Sivakumar H., Erali R.A., Wajih N., Li W., Shen P., Levine E.A., Miller K.E., Skardal A, & Votanopoulos K.I. (2022). Patient-Specific Sarcoma Organoids for Personalized Translational Research: Unification of the Operating Room with Rare Cancer Research and Clinical Implications. Annals of surgical oncology, 29(12), 7354-7367.

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Intracellular Cytokine Analysis of TILs

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For the intracellular cytokine analysis of TILs, cells from dissociated tumors in the density range of 1×10⁵ to 1×10⁶ cells were cultured in a 96-well U-bottom plate containing 10 µg/mL of pembrolizumab (Selleckchem, Houston, Texas, USA), 10 µg/mL of ipilimumab (Selleckchem), soluble 50 ng/mL of anti-CD3 (BD Biosciences, San Jose, California, USA), and 50 ng/mL of anti-CD28 (BD Biosciences) for 24 hours at 37°C, followed by the addition of brefeldin A (BD Biosciences) and monensin (BD Biosciences) per the manufacturer’s protocol. To measure interferon (IFN)-γ and tumor necrosis factor (TNF) secretion levels by CD8⁺ TILs according to PD-1 expression on anti-CD3 and anti-CD28 stimulation, the pre-incubation step without brefeldin A and monensin for 24 hours was omitted. After 12 hours, the cells were harvested and stained. Intracellular cytokines were stained after surface staining, fixation, and permeabilization with Foxp3 fixation/permeabilization solution (Thermo Fisher Scientific).

Kim C.G., Kim G., Kim K.H., Park S., Shin S., Yeo D., Shim H.S., Yoon H.I., Park S.Y., Ha S.J, & Kim H.R. (2021). Distinct exhaustion features of T lymphocytes shape the tumor-immune microenvironment with therapeutic implication in patients with non-small-cell lung cancer. Journal for Immunotherapy of Cancer, 9(12), e002780.

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Organoid Drug Treatment Evaluation

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Organoids were subsequently treated after 7 days of culture with 100 nM of Pembrolizumab (A2002, Selleckchem, Houston, TX), Ipilimumab (A2001, Selleckchem, Houston, TX) or Nivolumab (A2005, Selleckchem, Houston, TX). This drug concentration corresponds to dosing recommendations for a 70 kg patient. Media was aspirated from the wells and drug solutions mixed in culture media were added to each well individually. Organoids remained in drug-containing media solution for 3 days prior to endpoint viability assessment.

Forsythe S.D., Erali R.A., Sasikumar S., Laney P., Shelkey E., D’Agostino R J.r., Miller L.D., Shen P., Levine E.A., Soker S, & Votanopoulos K.I. (2021). Organoid Platform in Preclinical Investigation of Personalized Immunotherapy Efficacy in Appendiceal Cancer: Feasibility Study. Clinical cancer research : an official journal of the American Association for Cancer Research, 27(18), 5141-5150.

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