Human PLC/PRF/5 (Alexander cells) hepatocellular carcinoma cell line was procured from the cell repository of the National Centre for Cell Science (NCCS), Pune, India. Dulbecco’s Modified Eagle Medium (DMEM) was purchased from Hi-media and fetal bovine serum (FBS) was purchased from Gibco. Penicillin–streptomycin, trypsin, Live/Dead-Viability/Cytotoxicity Assay Kit, Annexin-PI staining kit, Cell MaskTM (plasma membrane staining kit) and Hoechst were purchased from Invitrogen – Thermofisher Scientific.
Dulbecco s modified eagle medium
Manufactured by HiMedia
Sourced in India
Dulbecco's Modified Eagle Medium (DMEM) is a cell culture medium commonly used for the growth and maintenance of various cell lines. It provides a balanced salt solution, amino acids, vitamins, and other nutrients required for cell growth and proliferation. DMEM is formulated to support the culture of a wide range of mammalian cell types, including fibroblasts, epithelial cells, and various transformed cell lines.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by HiMedia (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Phosphate buffered saline (pbs), by HiMedia (2 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Live dead viability cytotoxicity assay kit, by Thermo Fisher Scientific (1 mentions)
Cellmasktm, by Thermo Fisher Scientific (1 mentions)
Hoechst, by Thermo Fisher Scientific (1 mentions)
Thiazolyl blue tetrazolium bromide, by Merck Group (1 mentions)
Trypsin edta solution, by HiMedia (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Penicillin, by HiMedia (1 mentions)
Streptomycin, by HiMedia (1 mentions)
Vincristine sulfate, by Merck Group (1 mentions)
Nitrocellulose membrane, by GE Healthcare (1 mentions)
Verapamil hydrochloride, by Merck Group (1 mentions)
Piperine, by Merck Group (1 mentions)
Colchicine, by HiMedia (1 mentions)
Anti tubulin antibody, by Santa Cruz Biotechnology (1 mentions)
Protease inhibitor cocktail, by Merck Group (1 mentions)
10 protocols using dulbecco s modified eagle medium
1
Hepatocellular Carcinoma Cell Line Culture
2
Cytotoxicity Evaluation of Cell Lines
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Carboxymethyl cellulose (CM Cellulose), Dulbecco’s modified eagle medium (DMEM), Fetal bovine serum (FBS), Penicillin, Streptomycin and Trypsin-EDTA Solution were purchased from Himedia (India). Thiazolyl Blue Tetrazolium Bromide (MTT), Dimethyl Sulfoxide (DMSO), Neural Red (NR), Formaldehyde, β-Nicotinamide adenine dinucleotide reduced dipotassium salt (β-NADH), Trypan Blue and Triton X-100 dye solution were purchased from Sigma-Aldrich (USA).
3
Investigating Multidrug Resistance Mechanisms
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Verapamil hydrochloride, piperine, vincristine sulfate, rhodamine 123 (Rho123), dimethyl sulfoxide (DMSO), RIPA buffer, and protease inhibitor cocktail were purchased from Sigma-Aldrich, USA. Colchicine, Dulbecco’s Modified Eagle Medium (DMEM), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), trypsin-ethylene diamine tetra acetic acid (Trypsin–EDTA) solution, penicillin–streptomycin solution, phosphate-buffered saline (PBS), acridine orange (AO), ethidium bromide (EB), 4′,6-Diamidino-2-Phenylindole, dihydrochloride (DAPI), Bradford reagent and fetal bovine serum (FBS) were obtained from HiMedia (Mumbai, India). Nitrocellulose membrane was purchased from GE healthcare, USA. Anti-P-gp (D-11), and anti-tubulin antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
4
Culturing HepG2 Liver Cancer Cells
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The human hepatocarcinoma HepG2 cell line obtained from National Center for Cell Science (India) was maintained in low-glucose Dulbecco’s Modified Eagle Medium (HiMedia, India) supplemented with 10% fetal bovine serum, 100 UI/mL penicillin, 100 μg/mL streptomycin and 25 μg/mL amphotericin B. Cells were grown at 37 °C and 5% CO2 under controlled humidity. Sub-culturing was performed at approximately 45–50 h intervals, and cell growth was monitored with an Olympus inverted microscope. Cell count was performed by trypan blue method using an automated cell counting system (Farscope B, Curiosis, Seoul, Republic of Kore).
5
Optimizing Enzyme Activity Assays
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Chemicals
procured from Sigma-Aldrich [chitosan, malondialdehyde (MDA), dimethyl-sulfoxide
(DMSO), MF, and acetylcholinesterase (AChE) from electric eel (type-VI-S)],
chemicals procured from Spectrochem [1-(3-dimethylaminopropyl)-3-ethyl
carbodiimide-hydrochloride (EDC·HCL)], chemicals procured from
SD Fine [Tween 80], chemicals procured from Thermo Fisher Scientific
Inc. [Dulbecco’s modified Eagle medium (DMEM), 10% fetal bovine
serum (FBS), and (3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium
bromide) (MTT)], and chemicals procured from Himedia Laboratories
[5,5′-dithiobis(2-nitrobenzoic)acid (DTNB) and acetylthiocholine
iodide (ATCl)] were used.
procured from Sigma-Aldrich [chitosan, malondialdehyde (MDA), dimethyl-sulfoxide
(DMSO), MF, and acetylcholinesterase (AChE) from electric eel (type-VI-S)],
chemicals procured from Spectrochem [1-(3-dimethylaminopropyl)-3-ethyl
carbodiimide-hydrochloride (EDC·HCL)], chemicals procured from
SD Fine [Tween 80], chemicals procured from Thermo Fisher Scientific
Inc. [Dulbecco’s modified Eagle medium (DMEM), 10% fetal bovine
serum (FBS), and (3-(4,5-dimethythiazol-2-yl)-2,5-diphenyltetrazolium
bromide) (MTT)], and chemicals procured from Himedia Laboratories
[5,5′-dithiobis(2-nitrobenzoic)acid (DTNB) and acetylthiocholine
iodide (ATCl)] were used.
6
Infection Dynamics in Cell Lines
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Human lung carcinoma cells (A549; ATCC) and human hepatocellular carcinoma cells (Huh7; ATCC) were cultured in Dulbecco’s modified Eagle medium (HiMedia), human monocytic cells (THP1; ATCC) were cultured in RPMI 1640 medium (HiMedia), African green monkey kidney cells (Vero; ATCC) were cultured in minimum essential medium (HiMedia), and Aedes albopictus mosquito cells (C6/36; ATCC) were cultured in L-15 medium (HiMedia) with 10% fetal bovine serum (FBS) (HyClone; SH30070). All media were additionally supplemented with 100 μg/ml penicillin-streptomycin and 2 mM l -glutamine.
To perform the in vitro experiments, A549 and Huh7 cells were infected by DENV serotype 2 at a multiplicity of infection (MOI) of 5, and THP1 cells were infected at an MOI of 3 along with mock-infected control. Experiments were done in triplicates. The cells were incubated in the serum-free medium when inoculated with the virus for 2 h; the medium was then replaced with a 2% FBS-containing medium.
To perform the in vitro experiments, A549 and Huh7 cells were infected by DENV serotype 2 at a multiplicity of infection (MOI) of 5, and THP1 cells were infected at an MOI of 3 along with mock-infected control. Experiments were done in triplicates. The cells were incubated in the serum-free medium when inoculated with the virus for 2 h; the medium was then replaced with a 2% FBS-containing medium.
7
Comprehensive Pancreatic Cell Assays
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Atropine, Carbachol, FAEE, Taurolithocholic acid disodium salt, Bovine serum albumin, and Collagenase IV from Clostridium histolyticum were obtained from Sigma-Aldrich (St. Louis, USA); Lipopolysaccharide from Escherichia was purchased from Sigma-Aldrich (Rehovot, Israel). The substrates BOC-Gln-Ala-Arg-7AMC and Z-Arg-Arg-7AMC hydrochloride were obtained from Sigma-Aldrich (Buchs, Switzerland), LDH cytotoxicity assay kit was obtained from G-Biosciences (St. Louis, USA). Hisep LSM 1077, Dulbeccos modified eagle medium, Penstrep, Phosphate buffered saline, Fetal bovine serum were purchased from Himedia laboratories (Mumbai, India). Cytometric bead array (CBA) human inflammatory cytokines kit was purchased from BD Biosciences, USA. Autozyme, an amylase quantification kit was purchased from Accurex Biomedicals (Thane, India); recombinant TNF-α was purchased from Peprotech (Rocky Hill, NJ USA). Polyclonal primary antibodies (anti IL-6, anti-TNF-α, anti-LC3, anti caspase-3) were purchased from Abcam (Cambridge, UK), while anti-amylase antibody was purchased from Santacruz (California, USA). All other chemicals were obtained either from Sigma-Aldrich (Germany, USA) or SDFCL Fine chemicals limited (Mumbai, India).
8
Engineered Constructs in Osteosarcoma Cells
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The human osteosarcoma cell line (U2OS) was procured from ATCC (Manassas, VA, USA). U2OS cells were grown in DMEM (Dulbecco's Modified Eagle Medium; Himedia, Mumbai, India) supplemented with 10% Fetal Bovine Serum (Himedia) along with 1% penicillin-streptomycin (Invitrogen, Waltham, MA, USA) in a humidified incubator in the presence of 5% CO 2 at 37 °C. pcDNA3-bio-myc-mCE was obtained from Addgene (#82475; Watertown, MA, USA). The pcDNA4TO-bio-myc-mCE construct was prepared by sub-cloning the bio-myc-mCE sequence into an empty pcDNA4TO vector using KpnI and ApaI restriction sites. pcDNA4TO-bio-myc-K294A was prepared from pcDNA4TO-bio-myc-mCE using In-Fusion Site Directed Mutagenesis kit (Takara Bio, San Jose, CA, USA). The HA-NCK1-M3 construct was a generous gift from L. Larose [52] , and was prepared as described in Chen et al. [53] . Cells were transfected with either vector control pcDNA4TO, pcDNA4TO-bio-myc-K294A, vector control pRK5, or pRK5HA-NCK1-M3 using Jetprime transfection reagent (Polyplus, Illkirch-Graffenstaden, France) according to the manufacturer's guidelines. Cells were harvested after 48 h post-transfection.
9
Comprehensive Analytical Techniques for Cellular Assessments
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2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6sulphonic acid) (ABTS), nitroblue tetrazolium (NBT), butylated hydroxyanisole (BHA), Folin-Ciocalteu (FC), ethylenediaminetetraacetic acid (EDTA), 2,6-dichlorophenolindophenol (DCPIP), p-iodonitrotetrazolium chloride (INT), nutrient broth (NB), Dulbecco's Modified Eagle Medium (DMEM), propidium iodide (PI), acridine orange (AO), ethidium bromide (EB), 4′,6-diamidino-2-phenylindole (DAPI) and 2′,7′-dichlorofluorescin diacetate (DCFDA), were procured from Himedia (Mumbai, India). Fetal bovine serum (FBS) and water-soluble tetrazolium (WST) were obtained from Takara Bio Inc, Japan, and Invitrogen, Carlsbad, CA, USA, respectively. High-performance thin-layer chromatography (HPLC) standards were used from Sigma chemicals Co., St. Louis, MO, USA. 5,5′,6,6′-tetrachloro1,1′,3,3′tetraethylbenzimidazolcarbocyanine iodide (JC-1) and APO-DIRECT™ Kit for Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay were used from BD Pharmingen, San Diego, CA, USA.
10
DENV-2 Replicon Assay Protocol
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Baby Hamster Kidney (BHK-21) cells were procured from National Centre for Cell Science, Pune, India. Dulbecco's modified Eagle medium (Himedia; catalogue#AT149), FBS (Life Technologies; catalogue#10439-016), L-Glutamine (Sigma; catalogue#G7513), Penicillin-Streptomycin (Sigma; catalogue#P4333), mycophenolic acid (Sigma; catalogue#M5255) an inhibitor of DENV-RNA replication, were procured from the supplier (Sigma-Aldrich®). The cells were cultured and maintained at 37 °C with 5% CO 2 incubator (Thermo Scientific™ Forma™ Steri-Cycle i160).
The pRS424 plasmid containing frame work of non-structural genes of DENV 2 (NGCstrain) was a kind gift from Prof. Padmanabhan's laboratory (Georgetown University, Washington DC, USA). The pRS424 plasmid containing coding regions (RLuc-IRES-Neo r -DENV2-NS) were cloned downstream of SP6 promoter. The gene order of replicon is 5′ UTR, N terminal coding sequence of capsid (C), Renilla Luciferase reporter (Rluc) with translational termination codon and internal ribosome entry site (IRES) for 5′-cap-independent translation of the downstream ORF that encodes a poly protein precursor C terminus E-NS1-NS2A, NS2B, NS4A, NS4B, NS5 followed by 3′ UTR.
The pRS424 plasmid containing frame work of non-structural genes of DENV 2 (NGCstrain) was a kind gift from Prof. Padmanabhan's laboratory (Georgetown University, Washington DC, USA). The pRS424 plasmid containing coding regions (RLuc-IRES-Neo r -DENV2-NS) were cloned downstream of SP6 promoter. The gene order of replicon is 5′ UTR, N terminal coding sequence of capsid (C), Renilla Luciferase reporter (Rluc) with translational termination codon and internal ribosome entry site (IRES) for 5′-cap-independent translation of the downstream ORF that encodes a poly protein precursor C terminus E-NS1-NS2A, NS2B, NS4A, NS4B, NS5 followed by 3′ UTR.
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