Ovalbumin was obtained from Sigma-Aldrich (St. Louis, MO, USA). Brefeldin A solution and the following fluorescence-labeled antibodies, mouse PE/Cy7-anti-CD3 antibody, FITC-anti-CD4 antibody, FITC-anti-CD8a antibody, Percp-anti-CD8a antibody, PE-anti-IL4 antibody and APC-anti-IFN-γ antibody, were purchased from BioLegend (San Diego, CA). LysoTracker Red DND-99, RPMI medium 1640 basic, fetal bovine serum and penicillin streptomycin were purchased from Thermo Fisher Scientific (MA, USA). Mouse cytokine ELISA kits were obtained from BioLegend (San Diego, CA, USA). Recombinant mouse granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4 were purchased from PeproTech (Rocky Hill, USA).
Rpmi medium 1640 basic
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
RPMI Medium 1640 basic is a widely used cell culture medium formulation. It provides a balanced salt solution and essential nutrients to support the growth and maintenance of various cell types in in vitro cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (30 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (12 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (9 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Sartorius (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Matrigel, by BD (2 mentions)
Thp 1, by American Type Culture Collection (2 mentions)
Tgf β1, by Merck Group (2 mentions)
Mrc 5, by American Type Culture Collection (2 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (2 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (2 mentions)
Newborn calf serum nbcs, by Thermo Fisher Scientific (2 mentions)
Cck 8, by Dojindo Laboratories (2 mentions)
Cell counting kit 8 (cck8), by Dojindo Laboratories (2 mentions)
Lysotracker red dnd 99, by Thermo Fisher Scientific (1 mentions)
Anti ifn γ apc antibody, by BioLegend (1 mentions)
Ovalbumin (ova), by Merck Group (1 mentions)
Fitc anti cd4 antibody, by BioLegend (1 mentions)
59 protocols using rpmi medium 1640 basic
1
Activation of Mouse T-Cell Subsets
2
Antioxidant and Immune Response Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
GRCP was provided by Korea Food Research Institute. Fetal bovine serum (FBS), lipopolysaccharide (LPS), concanavalin (ConA), and methyl phenazine sulfate (PMS) were purchased from Sigma-Aldrich Co. (St. Louis, Mo, USA). Methyl trichlorosilane (MTS) was purchased from American Promega Co. RPMI Medium 1640 basic was purchased from Thermo Fisher Scientific (China) Co. Ltd. Protein extraction kit, Vitamin C (VC), d -galactose, and Red Blood Cell Lysis Buffer were bought from Beijing Solarbio Technology Co. Ltd. The assay kits for Total antioxidant capacity(T-AOC), Superoxide Dismutase (SOD), glutathione peroxidase (GSH-Px), Catalase (CAT), and malondialdehyde (MDA) were the products of Nanjing Jiancheng Bioengineering Institute (Nanjing, China). The enzyme-linked immunosorbent assay (ELISA) kit of Interleukin 1β (IL-1β), Interleukin 2 (IL-2), Interleukin 4 (IL-4), Interleukin 6 (IL-6), lipofuscin (Lipo), Tumor necrosis factor α (TNF-α) and Transforming Growth factor β(TGF-β) were purchased from Shanghai Yuchun Bio-tech Inc (Shanghai, China). All the other chemicals and reagents used in this study were of analytical grade made in China unless otherwise indicated. The experimental water was distilled water prepared by our laboratory.
3
Matrigel-Coated Transwell Invasion Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Matrigel‐coated Transwell assays were performed as previously reported.30 HTR8/SVneo cells (8 × 104) were plated in transwell inserts (8.0 μm, Merck Millipore, Germany) precoated with 60 μl of Matrigel (BD Biosciences). After 4 h of incubation at 37°C, 200 μl of RPMI Medium 1640 basic (1×; Life Technologies) containing vehicle (0.1% DMSO), AICAR (Selleck), compound C (Selleck) or PEP was loaded into the upper chambers, while 500 μl of medium containing 10% FBS was added into the lower chambers. After incubation in 21% O2 or 1% O2 at 37°C for 24 h, the noninvading cells on the top of the inserts were scraped off using a cotton swab. The filter under the inserts with invaded cells attached was washed with cold PBS, and the cells were fixed with 4% paraformaldehyde, stained with crystal violet and photographed with an EVOS FL Auto microscope (Life Technologies). In each independent experiment, a scan of the entire field of view was obtained for a comprehensive description of each sample, and 10 fields of view at 200× magnification were randomly selected per sample for quantification.
4
Porcine Alveolar Macrophages and Cell Lines for PRRSV
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Primary and immortalized porcine alveolar macrophages (PAMs) were cultured in RPMI Medium 1640 basic (Life Technologies, Carlsbad, California, USA), supplied with 10% fetal bovine serum (FBS, Hyclone, Logan, Utah, USA). Marc-145 (a monkey kidney cell line), PK-15 (a porcine kidney cell line), PT-K75 (pig intranasal mucosal fibroblasts), and HEK-293T (human embryonic kidney) were cultured in Dulbecco’s minimum essential medium (Life Technologies, USA), also supplied with 8% FBS (Hyclone, USA), 100 U/mL penicillin, and 100 µg/mL streptomycin. The SPF pigs used in the experiments were sourced from Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences. The animal experiments conducted were approved by the Animal Care and Use Committee of Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences (Approval ID: 200720-01). All experiments were performed in accordance with the regulations and guidelines established by this committee. The PRRSV North American-like strain HuN4 (GenBank accession number EF635006) was grown and titrated in Marc-145 cells, as previously described, and was stored at −80 °C.
5
Synthesis of Gold Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Chloroauric acid were purchased from Aladdin Bio-Chem Technology Co., Ltd. (Shanghai, China). DNase/RNase-free distilled water (Dalian Meilun Biotechnology Co., Ltd, China) was used throughout the experiments. The oligonucleotides and primers were synthesized by Sangon Biotech (Shanghai) Co., Ltd. Dulbecco's Modified Eagle's Medium (DMEM) and RPMI Medium 1640 basic were obtained from GIBCO, Thermo Fisher Scientific Co., Ltd (Rockford, USA). Fetal bovine serum was provided by CellMax. Carbon fiber was purchased from Jilin Carbon Co., Ltd. All other chemicals not mentioned here were of analytical reagent grade.
6
Cytokine and Metabolic Stress Effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
NIH3T3 and L929 cells were obtained from Shanghai Zhong Qiao Xin Zhou Biotechnology Co.Ltd. The cells were cultured at 37 °C in a 5% CO2 atmosphere using RPMI Medium 1640 basic (1640; Gibco, New York, NY, USA) supplemented with 10% fetal bovine serum (FBS, Biological Industries, Brisbane, BI, Australia), 1:100 penicillin/streptomycin, and 100 mM nonessential amino acids(Gibco, USA). Cells were passaged using a standard protocol with 0.25% trypsin (Sigma, Saint Louis, MO, USA) and seeded 12 h before experiments. The cells were seeded in 6-well culture plates at a confluence of 50%. The cells were stimulated with 25 ng/mL IL11 (GenScript, Nanjing, China) and/or 50 µM L-KYN (Macklin, Shanghai, China) diluted with RPMI 1640 for 2 h. For hypoxic conditions, the cells were grown in a three-gas incubator (SANYO) with 1% oxygen, 5% carbon dioxide, and 94% nitrogen. For glucose deprivation conditions, PRMI-1640 medium without glucose (Gibco, USA) was used.
7
Breast Cancer Cell Line Cytotoxicity Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gemcitabine hydrochloride, MW 299.66, β-glucan, (hydroxypropyl)methyl cellulose (HPMC), ethylcellulose (with 48% ethoxy content) (EC), acetic acid, 3-(4, 5-dimethyl-thiazol-2-yl)-2, and 5-diphenyl tetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St. Louise, MO, USA). Methylcellulose LR was purchased from Chem-Supply (Gillman, South Australia). Chitosan (MW 150,000, DD 70%), was purchased from Comwin Fine Chemicals Co. (Changzhou, China). Methylcellulose (MC) was purchased from ICN Biomedical, Inc. (Santa Ana, CA, USA). Polypropylene glycol was purchased from Midwest Pharmaceutics (Hawke’s Bay, New Zealand). Dimethyl sulfoxide (DMSO) was purchased from Labpartner (Shanghai, China). The 4T1breast cancer cell line was purchased from the American Type Culture Collection (ATCC; Rockville, MD, USA). The Roswell Park Memorial Institute (RPMI) medium 1640 basic was purchased from Gibco (Grand Island, NE, USA), and fetal calf serum was purchased from Hyclone (Logan, UT, USA). Penicillin–streptomycin–glutamine and nonessential amino acids were purchased from Life Technology (Grand Island, NY, USA). All other chemicals used were of reagent grade. Milli-Q water was obtained through reverse osmosis using a Millipak® system (Millipore, Burlington, MA, USA, 0.22 µm).
8
Macrophage and Fibroblast Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mouse macrophages (RAW 264.7) and fibroblasts (NIH/3T3), as well as the human monocytes (THP-1) and fibroblasts (MRC-5) were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). All cell lines were maintained in Dulbecco’s modified Eagle’s medium (DMEM, Life Technologies/Gibco, Grand Island, NY) or RPMI Medium 1640 basic (1640, Life Technologies/Gibco, Grand Island, NY) containing 10% fetal calf serum (FCS, Life Technologies/Gibco, Grand Island, NY), 100 U/ml penicillin and 100 μg/ml streptomycin (Life Technologies/Gibco, Gaithersburg, MD). The cell lines were cultured in a humidified atmosphere containing 5% CO2 at 37 °C. The human macrophages were acquired by using THP-1 cells differentiated with 100 nM PMA (Phorbol-12-myristate-13- acetate, Sigma, St. Louis, MO, USA) for 48–72 h.
For western blot or qRT-PCR analysis, macrophages and fibroblasts were plated (2 × 105 cells) into 6-well plates overnight. The macrophages were treated with 100 μg/ml silica for 12 h. The fibroblasts were treated with 1 ng/ml TGF-β1 (Sigma-Aldrich) for 48 h, and the total RNA or protein was prepared according to the experimental instructions.
For western blot or qRT-PCR analysis, macrophages and fibroblasts were plated (2 × 105 cells) into 6-well plates overnight. The macrophages were treated with 100 μg/ml silica for 12 h. The fibroblasts were treated with 1 ng/ml TGF-β1 (Sigma-Aldrich) for 48 h, and the total RNA or protein was prepared according to the experimental instructions.
9
H1299 Cell Culture Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H1299 cells (American Type Culture Collection) were cultured in RPMI Medium 1640 basic (Gibco, C11875500BT) supplemented with 10% fetal bovine serum (Gibco, 10091148), 1% penicillin and streptomycin (Gibco, 10378016) under cell culture conditions (37 °C, 5% CO2).
10
Cell Culture Protocols for Cancer Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
H22, HepG2, and RAW264.7 cells were obtained from the China Center for Type Culture Collection at Wuhan University. H22 cells were cultured in RPMI medium 1640 basic (Gibco) supplemented with 10% FBS (SeraPro, S601S-500) and antibiotics (100 mg/L streptomycin and 1 × 105 U/L penicillin) in a humidified atmosphere (5% CO2, 37 °C). Both HepG2 and RAW264.7 cell lines were maintained in DMEM containing 10% FBS and antibiotics in a humidified atmosphere.
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!