Human colon cancer cell lines: DLD-1 (ATCC: CCL-221TM),) were used. mRNA expression analysis was performed on DLD-1 in which LOX-1 mRNA was stably downmodulated (LOX-1#5) as previously reported in29 (link). DLD-1 scramble cells were used as control. Human colon cancer cells were grown in RPMI-1640 (Gibco, Life Technologies Corporation, Carlsbad, CA, USA) supplemented with 15% fetal bovine serum (FBS) (Euroclone, Milan, IT), Glutamine (Euroclone, Milan, IT), non-essential Amino Acids (Gibco, Life Technologies Corporation, Carlsbad, CA, USA), Penicillin–Streptomycin (Gibco, Life Technologies Corporation, Carlsbad, CA, USA).Cells were seeded at the density of 50,000 cells/cm2 and incubated at 37 °C, 5% CO2 for 24 h. Afterwards bevacizumab (Sigma-Aldrich S.r.l., Milan, Italy) was added at the concentration of 125 and 250 µg for 24 h. After this period, cells were trypsinized and collected. The experiment was performed in triplicate.
Bevacizumab
Manufactured by Merck Group
Sourced in France, Germany
Bevacizumab is a recombinant humanized monoclonal antibody that binds to and inhibits the biological activity of human vascular endothelial growth factor (VEGF). It is used as a laboratory reagent for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Bevacizumab, by Roche (3 mentions)
Glutamine, by Euroclone (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Euroclone (1 mentions)
Sodium dihydrogen phosphate monohydrate, by Merck Group (1 mentions)
U87 cell line, by American Type Culture Collection (1 mentions)
Disodium hydrogen phosphate dihydrate, by Merck Group (1 mentions)
Penicillin, by Lonza (1 mentions)
Streptomycin, by Lonza (1 mentions)
Sephadex g 50, by Merck Group (1 mentions)
111in chloride, by PerkinElmer (1 mentions)
Sodium bicarbonate, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Sodium citrate, by Merck Group (1 mentions)
Sunitinib, by Merck Group (1 mentions)
Dasatinib, by Merck Group (1 mentions)
Gemcitabine, by Merck Group (1 mentions)
Oxaliplatin, by Merck Group (1 mentions)
10 protocols using bevacizumab
1
Bevacizumab Alters LOX-1 mRNA in Colon Cancer
2
Optimizing U87 Cell Culture Conditions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The U87 cell line (ATCC, Rockville, USA) was maintained in Eagle's minimal essential medium (EMEM) with 10% fetal calf serum, 2 mM L-glutamine, 100 U/mL Penicillin and 100 µg/mL Streptomycin (Lonza, Verviers, Belgium).
Bevacizumab (Roche, Paris, France) was diluted with culture medium to working concentrations before use. SU1498 (EMD Chemicals, San Diego, USA), a selective VEGFR2 tyrosine kinase inhibitor [16] (link), was prepared as a stock solution of 30 mM in DMSO, then diluted with culture medium to working concentrations before use. As a control to Bevacizumab, a stock solution containing the corresponding excipient was prepared with 60 mg/mL α,α trehalose dihydrate; 5.8 mg/mL sodium dihydrogen phosphate monohydrate and 1.5 mg di-sodium hydrogen phosphate dihydrate (all from Sigma Aldrich, Saint-Quentin Fallavier, France).
Bevacizumab (Roche, Paris, France) was diluted with culture medium to working concentrations before use. SU1498 (EMD Chemicals, San Diego, USA), a selective VEGFR2 tyrosine kinase inhibitor [16] (link), was prepared as a stock solution of 30 mM in DMSO, then diluted with culture medium to working concentrations before use. As a control to Bevacizumab, a stock solution containing the corresponding excipient was prepared with 60 mg/mL α,α trehalose dihydrate; 5.8 mg/mL sodium dihydrogen phosphate monohydrate and 1.5 mg di-sodium hydrogen phosphate dihydrate (all from Sigma Aldrich, Saint-Quentin Fallavier, France).
3
Effects of VEGF and Bevacizumab on Neurons
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The effects of VEGF and bevacizumab on neurons and glial cells were investigated by addition of 0.1 μg/mL VEGF‐165 (SRP4365; Sigma‐Aldrich), 0.25 mg/mL bevacizumab (Avastin, B7106; Roche, Grenzach‐Wyhlen, Germany), or combination of VEGF with bevacizumab to the nutrient medium for periods of 10, 20, and 30 days starting at day 4 in culture. The role of VEGF receptors was experimentally examined via the use of the inhibitor axitinib (S1005; Selleckchem, Houston, TX, USA). This was used at a final concentration of 10 μM. The medium was changed twice a week.
4
Radiolabeling of Bevacizumab with Indium-111
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bevacizumab (25 mg/mL, Roche, USA) buffered with sodium bicarbonate (0.1 M, pH 8.2, Sigma-Aldrich) was reacted with 7-fold molar excess of 2-(4-isothiocyanatobenzyl) diethylenetriamine pentaacetic acid (DTPA, Macrocyclics, Dallas, USA) dissolved in anhydrous dimethyl sulfoxide (DMSO, Sigma-Aldrich) for 45 min at room temperature. Unreacted DTPA was removed by gel-filtration chromatography using a G-50 Sephadex (Sigma-Aldrich) column. Purified DTPA-Bevacizumab was buffer-exchanged into sodium citrate (0.1 M, pH 5.0, Sigma-Aldrich) and incubated with 111In chloride (1–2 MBq/μg) (PerkinElmer) for 1 h at room temperature. Radiolabelling yield was measured using instant thin layer chromatography (ITLC) in sodium citrate (0.1 M, pH 5.0) and was always >95%.
5
Synthesis and Characterization of Anti-API-5 Peptide
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The Anti-API-5 Peptide was synthesized by Proteogenix (Strasbourg, France) and was >95% pure, as determined by HPLC and mass spectrographic analysis. The sequence of the API-5 peptide corresponds to the Leucine-Zipper subdomain of API-5 with the Antennapedia sequence included in the first 15 amino acids, as described previously (Supplementary Figure 1A). The specificity of the peptide has been described previously [7 (link), 8 (link)]. The sequence is: RQIKIWFQNRRMKWKKAKLNAEKLKDFKIRLQYFARGLQVYIRQLRALQGKT.
All other drugs, including cisplatin, paclitaxel, epirubicin, sunitinib, dasatinib, everolimus, oxaliplatin, bevacizumab, and gemcitabine, were purchased from Sigma (France).
All other drugs, including cisplatin, paclitaxel, epirubicin, sunitinib, dasatinib, everolimus, oxaliplatin, bevacizumab, and gemcitabine, were purchased from Sigma (France).
6
Anti-cancer Effects of miR-148a
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To study the anticancer effects of miR-148a, we first treated the HCT116, HCT116-miR-148a, HT29, and HT29-miR-148a cells with various concentrations (0 mg/ml, 0.125 mg/ml, 0.25 mg/ml, 0.5 mg/ml, and 1 mg/ml) of bevacizumab for 24, 48, and 72 h, respectively, and evaluated cell viability by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay kit (Sigma-Aldrich, St. Louis, MO, USA). We seeded the cells (5 × 103 cells/well) into a 96-well culture plate and incubated the plate overnight at 37° C before treatment with the appropriate dose of bevacizumab (Avastin) (0, 0.125, 0.25, 0.5, or 1 mg/mL) (Roche). After 24-, 48-, and 72-h incubation, 20 μL of 5 mg/mL MTT was added to each well and incubated at 37° C for 2 h. The medium was replaced with 100 μL of dimethyl sulfoxide to dissolve the precipitate. Thereafter, absorbance was measured at 570 nm on a 96-well microplate reader (BioTeK Instruments, Winooski, VT, USA).
7
Targeting VEGF-A and PPARγ in A549 and H520 cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
VEGF-A inhibitor (bevacizumab, 2.5 μM) and PPARγ inhibitor, GW9662 (20 μM, Sigma-Aldrich, St. Louis, MO) were used to treat A549 and H520 cells for 72 h and harvested for further analysis.
8
Radiolabeling of Bevacizumab with 52Mn
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
bevacizumab (Avastin®; Roche Pharma AG, Grenzach-Wyhlen, Germany) was derivatized with a fivefold excess of 2,2′,2″-(10-(1-carboxy-4-((4-isothiocyanatobenzyl)amino)-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (p-NCS-Bn-DOTA-GA; Chematech) in NaHCO3 buffer (pH 8.2) at room temperature. DOTAGA-bevacizumab was purified by ultrafiltration on an Amicon Ultra (Merck KGaA, Darmstadt, Germany), 0.5 mL, 30 kDa centrifugal filter (Millipore). Radiolabeling was implemented at pH 6 for 15 min at room temperature. The required pH value was adjusted by sodium-acetate (0.04 M) buffer and sodium hydroxy. The efficiency of radiolabeling was followed by Raytest miniGita Star radio-thin layer chromatography scanner. 3 µL of the reaction mixture was dropped onto a glass macrofibre chromatography paper impregnated with silica gel (iTLC-SG) strip and developed in 0.1 M sodium citrate solution. [52Mn]Mn-DOTAGA-bevacizumab was remained near the start point (Rf = 0.1–0.2), whereas 52MnII as citrate complex was eluted with the solvent front (Rf = 0.8–1). The [52Mn]Mn-DOTAGA-bevacizumab product was used without further purification in experiments.
9
Bevacizumab and Pembrolizumab Combination in Solid Tumors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The primary objective of the phase Ib portion was determining MTD, the safety, and DLTs of the combination. The primary objective of the phase II portion was objective ORR, as measured by RECIST v1.1. Secondary objectives included PFS and OS.
Bevacizumab was sourced from a commercial supply, and pembrolizumab was provided by Merck & Co. Bevacizumab and pembrolizumab were infused intravenously over approximately 30 and 60 minutes, respectively, on day 1 of every 21-day cycle and administered 15-30 minutes apart. Treatment continued until disease progression, unacceptable toxicity, withdrawal of informed consent, or patient’s death.
The study definition of DLT is provided in the study protocol (Data Supplement). The MTD was defined as the dose level below the dose that induced DLTs in at least one third of the patients. If an MTD was not determined, the highest tested dose level (200 mg pembrolizumab and 15 mg/kg Bevacizumab) was defined as the recommended phase II dose.
Imaging of the chest, abdomen, and pelvis was performed every 6 weeks through cycle 9; thereafter, it was performed every 9 weeks. Response was assessed per the RECIST v1.118 (link) and immune-related RECIST.19 (link) Toxicities were graded using the National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE; version 4.0).
Bevacizumab was sourced from a commercial supply, and pembrolizumab was provided by Merck & Co. Bevacizumab and pembrolizumab were infused intravenously over approximately 30 and 60 minutes, respectively, on day 1 of every 21-day cycle and administered 15-30 minutes apart. Treatment continued until disease progression, unacceptable toxicity, withdrawal of informed consent, or patient’s death.
The study definition of DLT is provided in the study protocol (Data Supplement). The MTD was defined as the dose level below the dose that induced DLTs in at least one third of the patients. If an MTD was not determined, the highest tested dose level (200 mg pembrolizumab and 15 mg/kg Bevacizumab) was defined as the recommended phase II dose.
Imaging of the chest, abdomen, and pelvis was performed every 6 weeks through cycle 9; thereafter, it was performed every 9 weeks. Response was assessed per the RECIST v1.118 (link) and immune-related RECIST.19 (link) Toxicities were graded using the National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE; version 4.0).
10
Antibody Purification and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Honeywell, VWR Chemicals) and used without further purification. The protein ladder for SDS-PAGE analysis was the Precision Plus Protein Standard Dual Xtra (BioRad). Peptides were purchased from GeneCust and the antibodies bevacizumab (Bmab) and cetuximab (Cmab) were provided by Centre de lutte contre le Cancer Paul Strauss (France) and originally purchased from Merck KGaA and Roche laboratory, respectively. The initial buffer solution of the antibodies Cmab and Bmab was changed to PBS using illustra NAP-10 column (GE Healthcare). Antibodies used for the western blot analysis were purchased from Cell Signaling.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!