N3-PEG2000-ACA was purchased from MeloPEG (Shengzhen, China). 1,4-butanediol diacrylate (HDD), 4,4’-trimethyldipiperidine (TDP), tris (3-hydroxypropyltriazolyl methyl) amine (THPTA), fructose-6-phosphate (F6P), IR-780 were purchased from Aladdin (Shanghai, China). BAY-876 and UDP-GlcNAc were purchased from MedChemExpress (Shanghai, China). Annexin V-FITC apoptosis detection kit, DAPI, and TUNEL detection kit were purchased from Beyotime (Shanghai, China). ELISA kit of IFN-γ, TNF-α, CXCL-10 and granzyme were purchased from ABclonal (Wuhan, China). Fructose-6-Phosphate (F6P) Assay Kit was purchased from Sigma-Aldrich. Red blood cell lysis buffer and Glycerol anhydrous were obtained from Solarbio (Beijing, China). Mouse-PD1 protein, Mouse-biotin-PD-L1 and Mouse-biotin-CTLA-4 were purchased from ACROBiosystems (Beijing, China). Streptavidin-coated magnetic beads were purchased from BEAVER (Suzhou, China). DNA was purchased from Sangon (Shanghai, China), and the corresponding sequence information was provided in Supplementary Table 1 . siNC and siPD-L1 were purchased from Biosyntech (Suzhou, China), and the corresponding sequence information was provided in Supplementary Table 2 . The information of antibodies was provided in Supplementary Tables 3 and 4 .
Bay 876
Manufactured by MedChemExpress
Sourced in United States, China
BAY-876 is a chemical compound used as a laboratory tool. It functions as a selective inhibitor, targeting a specific biological mechanism. The core purpose of BAY-876 is to facilitate research and study within controlled laboratory settings.
Automatically generated - may contain errors
Lab products found in correlation
Annexin 5 fitc apoptosis detection kit, by Beyotime (1 mentions)
Tunel detection kit, by Beyotime (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Udp glcnac, by MedChemExpress (1 mentions)
Cxcl 10, by ABclonal (1 mentions)
Red blood cell lysis buffer, by Solarbio (1 mentions)
Tnf α, by ABclonal (1 mentions)
Doxorubicin, by Cayman Chemical (1 mentions)
Hrp conjugated anti rabbit igg, by Wuhan Servicebio Technology (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Wuhan Servicebio Technology (1 mentions)
Anti pd l1 antibody, by Wuhan Servicebio Technology (1 mentions)
Anti cd11b percp cy5.5 antibody, by Liankebio (1 mentions)
Celltiter glo luminescent cell viability assay kit, by Promega (1 mentions)
Penicillin, by Nacalai Tesque (1 mentions)
Streptomycin, by Nacalai Tesque (1 mentions)
Minimum essential medium (mem), by Nacalai Tesque (1 mentions)
D glucose, by Nacalai Tesque (1 mentions)
β actin 3700, by Cell Signaling Technology (1 mentions)
Ferroorange, by Dojindo Laboratories (1 mentions)
Liperfluo, by Dojindo Laboratories (1 mentions)
8 protocols using bay 876
1
Immunosuppressive Pathway Targeting Protocol
2
Targeting Ccne1+ Tumors with BAY-876 and Doxorubicin
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Ccne1+ tumor bearing mice were treated with 5 mg/kg BAY-876 (MedChemExpress) by oral gavage daily for 9 days beginning when the tumor volume reached 200 mm3. Doxorubicin (Cayman Chemical) was administered i.p. at 6 mg/kg every 3 days for 9 days beginning when the tumor volume reached 200 mm3.
3
Comprehensive Protocol for Tumor Immunotherapy
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BAY‐876 and BMS‐1 were purchased from MedChemExpress (MCE, China). L‐Ala‐NCA (CAS: 2224‐52‐4) and mPEG45‐NH2 (CAS: 80506‐64‐5) were purchased from Aladdin (China). LA assay kits were obtained by Jianchengbio (China). CCK‐8 assays, lysed RIPA buffer, PMSF, DAPI anti‐β‐tubulin antibody, antiIgG‐FITC antibody, anti‐IgG‐Cy3 antibody, and anti‐GLUT1 antibody, anti‐PD‐L1 antibody, HRP‐conjugated anti‐rabbit IgG and antimouse IgG were purchased from Servicebio (China). Murine IFN‐γ, TNF‐α, and TGF‐β ELISA kits, a cytokine of murine IFN‐γ were provided by Neobioscience (Shenzhen, China). anti‐PD‐L1‐APC antibody, anti‐CD80‐PE antibody, anti‐MHCII‐FITC antibody, anti‐CD45‐PE antibody, anti‐CD45‐FITC antibody, anti‐CD11b‐PerCP‐Cy5.5 antibody, anti‐CD3‐PerCP‐Cy5.5 antibody, anti‐CD206‐APC antibody, anti‐CD4‐FITC antibody, anti‐Foxp3‐APC antibody, anti‐CD11b‐PerCP‐Cy5.5 antibody, anti‐IFN‐γ‐APC antibody were purchased from Liankebio (China). anti‐CD206‐APC antibody was provided by Elabscience (China).
4
Measuring Cell Viability with CellTiter-Glo
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CellTiter-Glo™ Luminescent Cell Viability Assay Kit (Cat#G7570, Promega, Wisconsin, WI, USA) was used to measure cell viability according to manufacturer’s instructions. In brief, 3 x 104 cells were seeded in white-wall 96-well plates, 24 h after incubation; cells were treated with indicated reagents (H2O2 with or without 2-DG (Cat#HY-13966, MedChemExpress, New Jersey, NJ, USA)/Bay876 (Cat#HY-100017, MedChemExpress)) for 3 h. Then, 100 μL CellTiter-Glo® reagent was added to the experimental wells, incubated at room temperature for 10 min, and then luminescence was recorded with a luminometer.
5
Calu-3 Cell Culture and Glucose Stimulation
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Calu-3, a human submucosal gland cell line derived from bronchial adenocarcinoma, was purchased from Elabscience Biotechnology Inc. (USA). Calu-3 cells were cultured at 37 °C with 5% CO2 in MEM (Nacalai Tesque, Inc., Kyoto, Japan) with 10% fetal bovine serum (Hyclone, Logan, UT, USA), 100 IU/mL penicillin, and 100 μg/mL streptomycin (Nacalai Tesque, Inc.). The cells were stimulated with different D-glucose concentrations as follows: 100 mg/dL D-glucose (Nacalai Tesque, Inc.) in MEM (NG) and 1000 mg/dL D-glucose in MEM (HG). The medium was changed every alternate day. BAY-876 (Medchemexpress, Monmouth Junction, NJ, USA) was used to inhibit GLUT1.
6
Ferroptosis Regulation Mechanism Exploration
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Sodium selenite (Se), methylseleninic acid (MSeA), dihydroethidium (DHE), 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA), glutamine (Gln), glucose and manganese (III) tetrakis (1-methyl-4-pyridyl) porphyrin (MnTMPyP) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). BAY-876, deferoxamine mesylate (DFO) and ferrostatin-1 (Fer-1) were purchased from MedChem Express (Monmouth Junction, NJ, USA). Monosodium glutamate (MSG), salicylazosulfapyridine (SAS) and 2-deoxy-D-glcose (2-DG) were purchased from Aladdin (Shanghai, China). FerroOrange and Liperfluo were purchased from DOJINDO (Kyushu, Japan). Antibodies specific for SLC7A11 (12691), SLC7A11 (98051) and β-actin (3700) were purchased from Cell Signaling Technology (Beverly, MA, USA). The secondary antibodies: Horseradish peroxidase-linked Goat Anti-Rabbit IgG and Horseradish peroxidase-linked Goat Anti-Mouse IgG were obtained from MBL International Corporation (Beijing, China). SLC7A11 small interfering RNA (siRNA) and nontargeting siRNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
7
Evaluating Cytotoxicity of Pharmaceutical Compounds
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To determine the IC50 of sunitinib (MedChemExpress; cat. no. HY-10255A), BAY-876 (MedChemExpress; cat. no. HY-100017) and NaSH on 786-O viability, cells were seeded in a 96-well culture plate (4x103 cell/well) and incubated at 37˚C in a humidified incubator with 5% CO2. After 24 h, the cells were treated with various concentrations of sunitinib (range, 1-100 µM), BAY-876 (range, 10-1,000 µM) and NaSH (range, 10-103 µM) in quadruplicate for each concentration. The plates were incubated for 24, 48 and 72 h at 37˚C with 5% CO2. For BAY-876 another plate was incubated for 96 h. After each incubation interval, the culture media of the plate wells were changed with new media and then the cell cytotoxicity was measured by using Cell Counting Kit-8 (CCK-8; MedChemExpress; cat. no. HY-K0301) (24 (link)) according to the manufacturer's instruction. Briefly, 10 µM of the CCK-8 reagent was added to each well, incubated for 2-4 h at 37˚C and then the absorbance was measured at 540 nm by using an ELIZA reader (BioTek Instruments, Inc.). Each experiment was carried out three times.
8
Preclinical Evaluation of BAY-876 in PDX Models
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All animal experiments were reviewed and approved by the Animal Care Committee at the University Health Network in Toronto. For in vivo dosing experiments, BAY-876 was purchased from MedChemExpress (Cat. no. HY-100017). PDX tumors were subcutaneously implanted into SCID mice following standard procedure. BAY-876 was prepared at 10% NMP, 90% PEG300, and administered to the mice when tumors reach 100 mm3 in volume. The treatments were performed once daily by oral gavage for a total of 30 days. Body weights and tumor growth were measured once a week over the course of treatment until the endpoint was reached.
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