ATF3+/+ and ATF3-/- MEFs were obtained from Dr. Jim Dimitroulakos (Ottawa Hospital Research Institute) [12 (link)] and grown in DMEM High Glucose media (Hyclone, San Angelo) supplemented with 10% fetal calf serum (FBS) (Gibco, ThermoFisher Scientific, Ottawa) and 90 units/ml penicillin, 90 ug/ml streptomycin (Hyclone). HCT116 and HeLa cells were obtained from the American Tissue Type Collection (Cat #: CCL-247 and CCL-2, Manassas, VA) grown in McCoys and DMEM media (Hyclone), respectively, supplemented with a 3:1 mixture of Newborn Calf Serum (NBCS) (Gibco, Thermo Fisher Scientific) and Fetal Calf Serum (FBS) (Gibco, ThermoFisher Scientific), and 90 units/ml penicillin, 90 ug/ml streptomycin (Hyclone). Cell were seeded at a density of 500,000 cells/6 cm or 25,000 cells per well of a 96-well plate, 24 hours prior to treatment, depending on the assay. Cells were treated with up to 30 μM IGG (Calbiochem), up to 25nM PB (ChemCruz), 200nM actinomycin D (act D) (Sigma-Aldrich, St. Louis, MO), an equivalent volume of dimethyl sulfoxide (DMSO) as the vehicle control for IGG or left untreated.
Newborn calf serum nbcs
Manufactured by Thermo Fisher Scientific
Sourced in United States, China, Italy
Newborn calf serum (NBCS) is a biological substance derived from the blood of newborn calves. It is commonly used as a supplement in cell culture media to support the growth and proliferation of various cell types in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (6 mentions)
Penicillin, by Thermo Fisher Scientific (4 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (3 mentions)
Rpmi medium 1640 basic, by Thermo Fisher Scientific (2 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin p s, by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
M199 medium, by Thermo Fisher Scientific (2 mentions)
Collagenase treatment, by Merck Group (2 mentions)
Antibiotic antimycotic, by Merck Group (2 mentions)
L glutamine, by Merck Group (2 mentions)
Actinomycin d actd, by Merck Group (1 mentions)
Anti cd3 fitc, by Thermo Fisher Scientific (1 mentions)
Anti cd45 pe cy7, by Thermo Fisher Scientific (1 mentions)
Bms202, by Selleck Chemicals (1 mentions)
Hematoxylin and eosin h e staining solution, by Merck Group (1 mentions)
21 protocols using newborn calf serum nbcs
1
Cell Culture and Treatment Conditions
2
Synthesis of amino-terminated mPEG and reagents
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The amino-terminated poly (ethylene glycol) (mPEG45-NH2) was synthesized through the protocol described in our previous work (Chen et al., 2015 (link)). L-Leucine N-carboxyanhydride (L-Leu NCA) was obtained from Chengdu Enlai Biological Technology Co., Ltd. (Chengdu, China). Penicillin, streptomycin, trypsin-EDTA (0.05% trypsin and 0.02% EDTA) solution, RPMI 1640 medium, and new-born calf serum (NBCS) were bought from Gibco (Grand Island, NY, United States). Toluene, N,N-dimethylformamide (DMF; anhydrous), diethyl ether, elastase, and chymotrypsin were obtained from Aladdin Reagent Co., Ltd. (Shanghai, China). Hematoxylin and eosin (H&E) staining solution was purchased from Sigma-Aldrich (Shanghai, China). BMS202 was purchased from Selleck Chemicals (United States). PD-L1 antibody and Phospho-VEGF Receptor 2 antibody used for western blot (WB) were purchased from eBioscience (San Diego, CA, United States). PE-cy7-anti-CD45, FITC-anti-CD3, PerCP-cy5.5-anti-CD4, and Pacific Blue-anti-CD8 used for flow cytometry were purchased from eBioscience (San Diego, CA, United States). Enzyme-linked immunosorbent assay (ELISA) kits were purchased from Shanghai Lengton Bioscience Co., Ltd. (Shanghai, China). All the other chemicals were purchased from Beijing Chemical Industry Group Co., Ltd. (China).
3
Adipogenesis Protocol in 3T3-L1 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Dulbecco’s Modified Eagle’s Medium (DMEM), fetal bovine serum (FBS), and newborn calf serum (NBCS) were purchased from Gibco Life Technologies (Grand Island, NY, USA). Insulin, indomethacin, dexamethasone (DEX), 3-isobutyl-1-methylxanthine (IBMX), 3-(4,5)-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT), and Oil Red O (ORO) solution were purchased from Sigma-Aldrich (St. Louis, MO, USA). The 3T3-L1 preadipocytes were purchased from the Korea Cell Line Bank (KCLB, Seoul, Korea). For the Western blot analysis, the following antibodies were used: peroxisome proliferator-activated receptor gamma (PPARγ) and adiponectin (Abcam, Cambridge, MA, USA); CCAAT/enhancer binding protein alpha (C/EBPα), Insulin receptor β (IRβ), and anti-phosphorylated IRβ (p-IRβ) (Cell Signaling Technology, Beverly, MA, USA); and protein kinase B (AKT), phospho-AKT (p-AKT), and β-actin (Santa Cruz Biotechnology, Dallas, TX, USA).
4
Synthesis and Characterization of Heterocyclic Compounds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The imidazo[2,1-b][1,3,4]thiadiazole derivative IS1 and the indole derivatives IS2, IS3 and IS4 were synthesised at the Department of Pharmacy, University of Palermo, Italy, following the synthetic procedures previously described[39 (link),49 (link)]. The compounds were dissolved in dimethyl sulfoxide (DMSO), as reported previously[41 (link)]. Gemcitabine was kindly provided by Eli Lilly Corporation (Indianapolis, IN, USA) and dissolved in sterile water. Cell medium and newborn calf serum (NBCS) were from Gibco (Gaithersburg, MD, USA), while penicillin (50 IU mL-1) and streptomycin (50 µg mL-1) were from Lonza (Switzerland). Insulin-transferrin-selenium 100× was from Gibco (Grand Island, NY, USA) and PB was purchased from Cayman Chemical (Ann Arbor, MI, USA). All other chemicals were purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands).
5
Protocols for TMUV Infection in Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TMUV infections were performed using the following cell lines. C6/36 cells, an Aedes albopictus cell line gifted by Rui Luo, Huazhong Agricultural University, were cultured in RPMI medium 1640 basic (Gibco, Shanghai, China) supplemented with 10% fetal bovine serum (FBS) (Gibco, New York, USA) and 1% ciprofloxacin and incubated at 28°C with no additional CO2. BHK-21 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Shanghai, China) supplemented with 10% FBS and 1% ciprofloxacin and incubated at 37°C with 5% CO2. DEF cells were isolated from 9-day-old duck embryos, cultured in DMEM supplemented with 10% newborn calf serum (NBCS) (Gibco Shanghai China) and 1% ciprofloxacin, and incubated at 37°C with 5% CO2.
The duck-derived TMUV strain CQW1 (GenBank IDKM233707.1 ) was rescued from a DNA-based full-length infectious clone reported previously (53 (link)). The mosquito-derived TMUV strain MM 1775 (GenBank ID JX477685.2 ) was collected as mentioned above (24 (link)). All virus stocks were amplified in BHK-21 cells.
The duck-derived TMUV strain CQW1 (GenBank ID
6
Protocols for TMUV Infection in Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
TMUV infections were performed using the following cell lines. C6/36 cells, an Aedes albopictus cell line gifted by Rui Luo, Huazhong Agricultural University, were cultured in RPMI medium 1640 basic (Gibco, Shanghai, China) supplemented with 10% fetal bovine serum (FBS) (Gibco, New York, USA) and 1% ciprofloxacin and incubated at 28°C with no additional CO2. BHK-21 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Shanghai, China) supplemented with 10% FBS and 1% ciprofloxacin and incubated at 37°C with 5% CO2. DEF cells were isolated from 9-day-old duck embryos, cultured in DMEM supplemented with 10% newborn calf serum (NBCS) (Gibco Shanghai China) and 1% ciprofloxacin, and incubated at 37°C with 5% CO2.
The duck-derived TMUV strain CQW1 (GenBank IDKM233707.1 ) was rescued from a DNA-based full-length infectious clone reported previously (53 (link)). The mosquito-derived TMUV strain MM 1775 (GenBank ID JX477685.2 ) was collected as mentioned above (24 (link)). All virus stocks were amplified in BHK-21 cells.
The duck-derived TMUV strain CQW1 (GenBank ID
7
Differentiation of 3T3-L1 Preadipocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The 3T3-L1 cells were obtained from the American Type Culture Collection (ATCC # CL-173, Manassas, VA, USA). Insulin, isobutyl-methylxanthine (IBMX), dexamethasone (Dex), and Dulbecco’s modified Eagle’s medium (DMEM) were obtained from Sigma Aldrich (Oakville, ON, Canada). Fetal bovine serum (FBS), antibiotic-antimycotic (100×), and newborn calf serum (NBCS) were obtained from Gibco Life Technologies (Burlington, ON, Canada). Preadipocytes (3T3-L1) were grown to 70% confluency and were induced to differentiate by adding a differentiation cocktail (insulin (10 μg/mL), 1 μM Dex, and 0.5 mM IMBX) in DMEM containing 10% FBS along with 1% BM whey (both Q1 and Q4 from BMNO and BMO), as per previous publications [11 (link),20 (link),21 (link),22 (link)]. Control cells received only the differentiation cocktail. After 48 h, the media were changed to DMEM containing 10% FBS and insulin (10 μg/mL) along with 1% BM whey (Q1 and Q4) from both the BMNO and BMO groups. The media were replaced with DMEM containing 10% FBS + 1% BM whey every 48 h till day eight, when the cells were fully differentiated and harvested. The control cells received similar media at each stage of differentiation without BM whey.
8
HSV-1 KOS Viral Stock Generation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The HSV-1 KOS strain (ATCC-VR-1493) with 2x10 7 plaque forming units (pfu)/mL was used to prepare viral stocks after infection of Vero cells (ATCC-CCL-81). A confluent 75-cm 2 flask with Vero cells was infected with HSV-1 KOS at a multiplicity of infection (moi) of 0.01 pfu/cell. For this the virus was diluted in 3 mL of 199 medium (Gibco). After incubation for 2 h at 37°C the medium was aspirated and 3 mL of fresh DMEM (Gibco) supplemented with 5% New Born Calf Serum (NBCS; Gibco) was added to the flask. The virus was allowed to replicate for 3 days. Then the virus was released by freeze thawing following 3x sonication for 30 using a Misonix ultrasonic water bath at 100% amplitude. Aliquots of the virus were stored in liquid N 2 . The virus titer was determined by virus plaque assay.
9
Adipocyte Differentiation Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We purchased Dulbecco's modified Eagle medium (DMEM) from Gibco Life Technologies (#11965, Grand Island, NY); high glucose with l-glutamine, without sodium pyruvate, penicillin/streptomycin (P/S) from Invitrogen (#15140-122, Karlsruhe, Germany); new born calf serum (NBCS) from Gibco Life Technologies (#16170-078); fetal bovine serum (FBS) from Gibco Life Technologies (#10437-028); bovine insulin from Sigma-Aldrich (#I-5500, St. Louis, MO); isobutylmethyxanthine (IBMX) from Sigma-Aldrich (#I-7018); dexamethasone from Sigma-Aldrich (#D-4902); and dimethyl sulfoxide (DMSO) from ISC-Bioexpress (#0231, Kaysville, UT). Lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (#L2762), retinoic acid from Sigma-Aldrich (#R2625), and rosiglitazone from Enzo Life Sciences (#ALX-350-125-M100, Ann Arbor, MI).
The culture medium contained 1% P/S and 10% NBCS in DMEM. Two differentiation media were used. The first is termed differentiation medium I [DMEM containing 1% P/S and 10% FBS, bovine insulin (1:1,000), IBMX diluted in DMSO (1:1,000), and dexamethasone diluted in ethanol (1:10,000)]. Differentiation medium II was prepared with the same reagents as differentiation medium I; however, medium lacked the IBMX and dexamethasone. Differentiation was induced in confluent preadipocytes by differentiation medium I (d 0). Differentiation medium II was added on d 2 and replaced every 48 h.
The culture medium contained 1% P/S and 10% NBCS in DMEM. Two differentiation media were used. The first is termed differentiation medium I [DMEM containing 1% P/S and 10% FBS, bovine insulin (1:1,000), IBMX diluted in DMSO (1:1,000), and dexamethasone diluted in ethanol (1:10,000)]. Differentiation medium II was prepared with the same reagents as differentiation medium I; however, medium lacked the IBMX and dexamethasone. Differentiation was induced in confluent preadipocytes by differentiation medium I (d 0). Differentiation medium II was added on d 2 and replaced every 48 h.
10
Culturing 3T3-L1 and RAW 264.7 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
3T3-L1 preadipocyte cells and RAW 264.7 murine macrophage cells were obtained from the Food and Science Technology Department of National Pingtung Science Technology (NPUST, Pingtung, Taiwan). Both cell lines were maintained in Dulbecco's modified Eagle's medium (DMEM, Gibco, New York, USA) containing 1% penicillin/streptomycin (PS, 100 units of penicillin/mL and 100 pg streptomycin/mL, Gibco, New York, USA) and supplemented with 10% New-born Calf Serum (NBCS, Gibco, New York, USA) to culture the 3T3-L1 cells and 10% Fetal Bovine Serum (FBS, Gibco, New York, USA) to culture RAW 264.7 cells. The cells were incubated at 37°C in a humidified 5% CO 2 atmosphere and the medium was replaced every 2 days.
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!