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EuroClone hepes
Hepes, supplied by EuroClone, used in various techniques. Bioz Stars score: 94/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 19 article reviews
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hepes - by Bioz Stars, 2020-04
94/100 stars

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Cytometry:

Article Title: Mast Cells Respond to Candida albicans Infections and Modulate Macrophages Phagocytosis of the Fungus
Article Snippet: Precursor cells were cultivated in complete–IL-3 medium [RPMI 1,640 medium (Euroclone) supplemented with 20% FBS (Sigma Aldrich), 100 U·ml−1 penicillin, 100 μg·ml−1 Streptomycin, 2 mM Glutamine, 20 mM Hepes, non-essential amino acid, 1 mM Sodium Pyruvate (Euroclone), 50 mM β-mercaptoethanol (Sigma Aldrich), and 20 ng·ml−1 IL-3 (Peprotech)] at 37°C in 5% CO2 atmosphere. .. After 5 weeks, BMMCs differentiation was confirmed by flow cytometry by staining with fluorochrome-conjugated anti-FcεRIa (MAR-1) and anti-cKit (ACK2) antibodies (eBiosciences).

Real-time Polymerase Chain Reaction:

Article Title: 2-[4,5-Difluoro-2-(2-Fluorobenzoylamino)-Benzoylamino]Benzoic Acid, an Antiviral Compound with Activity against Acyclovir-Resistant Isolates of Herpes Simplex Virus Types 1 and 2
Article Snippet: These stocks were later quantified using quantitative PCR (qPCR) to determine the number of viral genomes per milliliter. .. The cells were grown in Dulbecco's modified Eagle medium (DMEM) (Sigma-Aldrich, St. Louis, MO) containing 0.75 mg/ml NaHCO3 , 20 mM HEPES (EuroClone, Milan, Italy), 1× PEST (100 IU/ml penicillin G, 100 μg/ml streptomycin sulfate; Gibco, Carlsbad, CA) and 5% fetal bovine serum (FBS) (Gibco) at 37°C.

Incubation:

Article Title: Facile Preparation of N-Glycosylated 10-Piperazinyl Artemisinin Derivatives and Evaluation of Their Antimalarial and Cytotoxic Activities
Article Snippet: Parasites were maintained at 5% hematocrit (human type A-positive red blood cells) in RPMI 1640 (EuroClone, Celbio) medium with the addition of 10% heat inactivate human serum, 20 mM Hepes and 2 mM glutamine (Euroclone). .. Asynchronous cultures of Pf with parasitaemia of 1–1.5% and 1% final hematocrit were aliquoted into 96-well flat-bottom microplates (COSTAR) with serial dilutions of test compounds and incubated for 72 h at 37 °C.

Article Title: Th17 Immunity in Children with Allergic Asthma and Rhinitis: A Pharmacological Approach
Article Snippet: Cell lines were grown in uncoated vented tissue culture flasks (Sarstedt, Numbrecht, Germany) and incubated at 37°C in a humidified 5% CO2 atmosphere. .. Cells were maintained in MEM (Gibco) supplemented with 10% foetal bovine serum (FBS) (Hyclone), 2mM glutamine (Euroclone), Hepes (Euroclone), gentamicin 250 μg/ml (Gibco) and 1% MEM non essential aminoacids (Gibco).

Article Title: Immunosuppression by monocytic myeloid-derived suppressor cells in patients with pancreatic ductal carcinoma is orchestrated by STAT3
Article Snippet: .. Cell cultures were incubated at 37 °C and 8% CO2 in arginine and glutamine–Free-RPMI (Biochrom AG, Berlin, Germany), supplemented with 2 mM L-glutamine (Euroclone, Milano, Italy), 150 μM arginine (Sigma-Aldrich, St. Louis, MO, USA), 10% FBS (Superior, Merck, Darmstadt, Germany), 10 U/ml penicillin and streptomycin (Euroclone, Milano, Italy), and 0.1 mM HEPES (Euroclone, Milano, Italy). .. At the end of the culture, cells were stained with PE-Cy7 conjugated anti-CD3 (UCHT1, eBioscience, Thermo Fisher Scientific, Waltham, MA, USA), and CellTrace signal of gated lymphocytes was analyzed.

Article Title: Induction of immunosuppressive functions and NF-κB by FLIP in monocytes
Article Snippet: PBMCs were recovered and washed in IMDM medium (Lonza, Visp, Switzerland), supplemented with 10% FBS (Euroclone, Milano, Italy), 100U/ml penicillin/streptomycin (Euroclone, Milano, Italy), β-mercaptoethanol (Sigma-Aldrich, Milan, Italy) and 10 mM HEPES (Euroclone, Milano, Italy). .. PBMCs were resuspended at a final concentration of 107 cells/ml in PBS and stained with 2,5 μM as final working concentration of CellTrace Violet stock solution (Thermo Fisher Scientific, Waltham, MA, USA), followed by 5 min’ incubation at 37 °C, protected from light.

Proliferation Assay:

Article Title: Immunosuppression by monocytic myeloid-derived suppressor cells in patients with pancreatic ductal carcinoma is orchestrated by STAT3
Article Snippet: Paragraph title: Human proliferation assay ... Cell cultures were incubated at 37 °C and 8% CO2 in arginine and glutamine–Free-RPMI (Biochrom AG, Berlin, Germany), supplemented with 2 mM L-glutamine (Euroclone, Milano, Italy), 150 μM arginine (Sigma-Aldrich, St. Louis, MO, USA), 10% FBS (Superior, Merck, Darmstadt, Germany), 10 U/ml penicillin and streptomycin (Euroclone, Milano, Italy), and 0.1 mM HEPES (Euroclone, Milano, Italy).

Article Title: Induction of immunosuppressive functions and NF-κB by FLIP in monocytes
Article Snippet: Paragraph title: Human proliferation assay ... PBMCs were recovered and washed in IMDM medium (Lonza, Visp, Switzerland), supplemented with 10% FBS (Euroclone, Milano, Italy), 100U/ml penicillin/streptomycin (Euroclone, Milano, Italy), β-mercaptoethanol (Sigma-Aldrich, Milan, Italy) and 10 mM HEPES (Euroclone, Milano, Italy).

Cell Culture:

Article Title: The Natural Compound Climacostol as a Prodrug Strategy Based on pH Activation for Efficient Delivery of Cytotoxic Small Agents
Article Snippet: .. Cell Cultures Murine melanoma B16-F10 cells, C2 C12 mouse myoblasts and NIH/3T3 mouse fibroblasts (Bizzozero et al., ; Cazzato et al., ; Assi et al., ; Basiricò et al., ; Cervia et al., ; Perrotta et al., ) were cultured in Iscove's or Modified Eagle Medium high glucose supplemented with 10% heat-inactivated fetal bovine serum, glutamine (2 mM), penicillin/streptavidin (100 U/ml), 1% Hepes 1 M (Euroclone), pH 7.2 (physiologic culture medium) (Cervia et al., , , ). ..

Article Title: Facile Preparation of N-Glycosylated 10-Piperazinyl Artemisinin Derivatives and Evaluation of Their Antimalarial and Cytotoxic Activities
Article Snippet: Cultures of Pf and In Vitro Antimalarial Assays The Pf strains D10 (CQ-sensitive), and W-2 (CQ-resistant) were cultured in vitro as described by Trager and Jensen with minor modifications [ ]. .. Parasites were maintained at 5% hematocrit (human type A-positive red blood cells) in RPMI 1640 (EuroClone, Celbio) medium with the addition of 10% heat inactivate human serum, 20 mM Hepes and 2 mM glutamine (Euroclone).

Article Title: The High Mobility Group A1 (HMGA1) Chromatin Architectural Factor Modulates Nuclear Stiffness in Breast Cancer Cells
Article Snippet: Paragraph title: 4.1. Cell Culture and Treatments ... MCF7 cells were grown in Dulbecco’s MEM Nutrient Mix F12 (1:1) with 25 mM HEPES (DME/F12-HEPES, Euroclone) supplemented with 2 mM L-glutamine, 10% tetracycline free fetal bovine serum (Tet-free FBS, Euroclone), penicillin (100 U/mL, Euroclone), streptomycin (100 μg/mL, Euroclone), and 1X MEM non-essential amino acids (Sigma Aldrich, St. Louis, MO, USA).

Expressing:

Article Title: The High Mobility Group A1 (HMGA1) Chromatin Architectural Factor Modulates Nuclear Stiffness in Breast Cancer Cells
Article Snippet: MCF7 cells were grown in Dulbecco’s MEM Nutrient Mix F12 (1:1) with 25 mM HEPES (DME/F12-HEPES, Euroclone) supplemented with 2 mM L-glutamine, 10% tetracycline free fetal bovine serum (Tet-free FBS, Euroclone), penicillin (100 U/mL, Euroclone), streptomycin (100 μg/mL, Euroclone), and 1X MEM non-essential amino acids (Sigma Aldrich, St. Louis, MO, USA). .. HMGA1 expression vector was obtained by subcloning the PCR products of the coding regions of human HMGA1a cDNA into the pcDNA3-HA plasmid expression vector (Invitrogen). pcDNA3-HA empty vector was used as a control.

Modification:

Article Title: The Natural Compound Climacostol as a Prodrug Strategy Based on pH Activation for Efficient Delivery of Cytotoxic Small Agents
Article Snippet: .. Cell Cultures Murine melanoma B16-F10 cells, C2 C12 mouse myoblasts and NIH/3T3 mouse fibroblasts (Bizzozero et al., ; Cazzato et al., ; Assi et al., ; Basiricò et al., ; Cervia et al., ; Perrotta et al., ) were cultured in Iscove's or Modified Eagle Medium high glucose supplemented with 10% heat-inactivated fetal bovine serum, glutamine (2 mM), penicillin/streptavidin (100 U/ml), 1% Hepes 1 M (Euroclone), pH 7.2 (physiologic culture medium) (Cervia et al., , , ). ..

Article Title: 2-[4,5-Difluoro-2-(2-Fluorobenzoylamino)-Benzoylamino]Benzoic Acid, an Antiviral Compound with Activity against Acyclovir-Resistant Isolates of Herpes Simplex Virus Types 1 and 2
Article Snippet: .. The cells were grown in Dulbecco's modified Eagle medium (DMEM) (Sigma-Aldrich, St. Louis, MO) containing 0.75 mg/ml NaHCO3 , 20 mM HEPES (EuroClone, Milan, Italy), 1× PEST (100 IU/ml penicillin G, 100 μg/ml streptomycin sulfate; Gibco, Carlsbad, CA) and 5% fetal bovine serum (FBS) (Gibco) at 37°C. .. Benzavir-1 and benzavir-2 were synthesized as described previously ( ).

Article Title: A Sialic Acid Binding Site in a Human Picornavirus
Article Snippet: .. Briefly, normal human conjunctival (NHC) cells were infected with CVA24v in serumfree medium containing Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich), HEPES (pH 7.4; EuroClone, Milan, Italy), and penicillin-streptomycin (PEST; Gibco, Carlsbad, CA) with gentle agitation for one hour at 37°C. .. Cells were washed with phosphate-buffered saline (PBS; Medicago AB, Uppsala, Sweden) to remove non-bound virions and then starved of methionine and cysteine in Met/Cys-free medium (Sigma-Aldrich).

Article Title: Splicing Mutations Impairing CDKL5 Expression and Activity Can be Efficiently Rescued by U1snRNA-Based Therapy
Article Snippet: Cell Cultures HEK293T and HeLa cells were maintained in DMEM (Dulbecco’s Modified Eagle Medium) (Sigma-Aldrich) supplemented with 10% FBS (Euroclone, Milan, Italy), 1% l -glutamine (Sigma-Aldrich) and 1% penicillin/streptomycin (Sigma-Aldrich) at 37 °C with 5% CO2 in T75 flasks. .. WT and c.2376+5G > A mutated Lymphoblastoid Cell Lines (LCLs) were maintained in RPMI medium-1640 (Roswell Park Memorial Institute) (Sigma-Aldrich), supplemented with 15% FBS, 1% L-glutamine, 1% penicillin/streptomycin, 2.5% HEPES (Euroclone) and 0.1% nystatin (Sigma-Aldrich) at 37 °C with 5% CO2 in 24-multiwell plates.

Western Blot:

Article Title: Splicing Mutations Impairing CDKL5 Expression and Activity Can be Efficiently Rescued by U1snRNA-Based Therapy
Article Snippet: Cells were seeded in 12-multiwell plates (Corning, Tewksbury, MA, USA) for RT-PCR and western blot or in 24-multiwell plates for immunofluorescence. .. WT and c.2376+5G > A mutated Lymphoblastoid Cell Lines (LCLs) were maintained in RPMI medium-1640 (Roswell Park Memorial Institute) (Sigma-Aldrich), supplemented with 15% FBS, 1% L-glutamine, 1% penicillin/streptomycin, 2.5% HEPES (Euroclone) and 0.1% nystatin (Sigma-Aldrich) at 37 °C with 5% CO2 in 24-multiwell plates.

Transformation Assay:

Article Title: Th17 Immunity in Children with Allergic Asthma and Rhinitis: A Pharmacological Approach
Article Snippet: Epithelial cell line culture 16-HBE (a differentiated SV-40 transformed bronchial epithelial cell) and human RPMI-2650 (squamous cell carcinoma of the nasal septum) (American Type Culture Collection, ATCC, Manassas, VA, USA) cell lines represent a valid in vitro model to evaluate the functional properties of bronchial and nasal epithelial cells in inflammation and repair processes , . .. Cells were maintained in MEM (Gibco) supplemented with 10% foetal bovine serum (FBS) (Hyclone), 2mM glutamine (Euroclone), Hepes (Euroclone), gentamicin 250 μg/ml (Gibco) and 1% MEM non essential aminoacids (Gibco).

Flow Cytometry:

Article Title: Mast Cells Respond to Candida albicans Infections and Modulate Macrophages Phagocytosis of the Fungus
Article Snippet: Precursor cells were cultivated in complete–IL-3 medium [RPMI 1,640 medium (Euroclone) supplemented with 20% FBS (Sigma Aldrich), 100 U·ml−1 penicillin, 100 μg·ml−1 Streptomycin, 2 mM Glutamine, 20 mM Hepes, non-essential amino acid, 1 mM Sodium Pyruvate (Euroclone), 50 mM β-mercaptoethanol (Sigma Aldrich), and 20 ng·ml−1 IL-3 (Peprotech)] at 37°C in 5% CO2 atmosphere. .. After 5 weeks, BMMCs differentiation was confirmed by flow cytometry by staining with fluorochrome-conjugated anti-FcεRIa (MAR-1) and anti-cKit (ACK2) antibodies (eBiosciences).

Infection:

Article Title: 2-[4,5-Difluoro-2-(2-Fluorobenzoylamino)-Benzoylamino]Benzoic Acid, an Antiviral Compound with Activity against Acyclovir-Resistant Isolates of Herpes Simplex Virus Types 1 and 2
Article Snippet: GMK cells were used throughout the study to assess HSV infection and the toxicities of the compounds. .. The cells were grown in Dulbecco's modified Eagle medium (DMEM) (Sigma-Aldrich, St. Louis, MO) containing 0.75 mg/ml NaHCO3 , 20 mM HEPES (EuroClone, Milan, Italy), 1× PEST (100 IU/ml penicillin G, 100 μg/ml streptomycin sulfate; Gibco, Carlsbad, CA) and 5% fetal bovine serum (FBS) (Gibco) at 37°C.

Article Title: A Sialic Acid Binding Site in a Human Picornavirus
Article Snippet: .. Briefly, normal human conjunctival (NHC) cells were infected with CVA24v in serumfree medium containing Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich), HEPES (pH 7.4; EuroClone, Milan, Italy), and penicillin-streptomycin (PEST; Gibco, Carlsbad, CA) with gentle agitation for one hour at 37°C. .. Cells were washed with phosphate-buffered saline (PBS; Medicago AB, Uppsala, Sweden) to remove non-bound virions and then starved of methionine and cysteine in Met/Cys-free medium (Sigma-Aldrich).

Generated:

Article Title: A Sialic Acid Binding Site in a Human Picornavirus
Article Snippet: 35 S-labeled CVA24v virions were generated as previously described . .. Briefly, normal human conjunctival (NHC) cells were infected with CVA24v in serumfree medium containing Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich), HEPES (pH 7.4; EuroClone, Milan, Italy), and penicillin-streptomycin (PEST; Gibco, Carlsbad, CA) with gentle agitation for one hour at 37°C.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Splicing Mutations Impairing CDKL5 Expression and Activity Can be Efficiently Rescued by U1snRNA-Based Therapy
Article Snippet: Cells were seeded in 12-multiwell plates (Corning, Tewksbury, MA, USA) for RT-PCR and western blot or in 24-multiwell plates for immunofluorescence. .. WT and c.2376+5G > A mutated Lymphoblastoid Cell Lines (LCLs) were maintained in RPMI medium-1640 (Roswell Park Memorial Institute) (Sigma-Aldrich), supplemented with 15% FBS, 1% L-glutamine, 1% penicillin/streptomycin, 2.5% HEPES (Euroclone) and 0.1% nystatin (Sigma-Aldrich) at 37 °C with 5% CO2 in 24-multiwell plates.

Immunofluorescence:

Article Title: Splicing Mutations Impairing CDKL5 Expression and Activity Can be Efficiently Rescued by U1snRNA-Based Therapy
Article Snippet: Cells were seeded in 12-multiwell plates (Corning, Tewksbury, MA, USA) for RT-PCR and western blot or in 24-multiwell plates for immunofluorescence. .. WT and c.2376+5G > A mutated Lymphoblastoid Cell Lines (LCLs) were maintained in RPMI medium-1640 (Roswell Park Memorial Institute) (Sigma-Aldrich), supplemented with 15% FBS, 1% L-glutamine, 1% penicillin/streptomycin, 2.5% HEPES (Euroclone) and 0.1% nystatin (Sigma-Aldrich) at 37 °C with 5% CO2 in 24-multiwell plates.

MTT Assay:

Article Title: Ellagitannins of the fruit rind of pomegranate (Punica granatum) antagonize in vitro the host inflammatory response mechanisms involved in the onset of malaria
Article Snippet: Cell cultures Human THP-1 monocytic leukaemia cells (ATCC, Teddington, UK) were grown in RPMI 1640 (Invitrogen s.r.l., Milano, Italy) supplemented with 100 units penicillin/ml, 100 μg streptomycin/ml, 10 mM HEPES, 1 mM Sodium Pyruvate, 0.05 mM beta-mercaptoethanol, and 10% heat-inactivated foetal calf serum (FCS, Euroclone, Milano, Italy). .. Cytotoxicity was evaluated by testing alterations of mitochondrial functionality as assessed by MTT test [ ].

Plaque Assay:

Article Title: 2-[4,5-Difluoro-2-(2-Fluorobenzoylamino)-Benzoylamino]Benzoic Acid, an Antiviral Compound with Activity against Acyclovir-Resistant Isolates of Herpes Simplex Virus Types 1 and 2
Article Snippet: Four of these isolates were also tested by plaque assay for resistance to foscarnet and were found to be foscarnet sensitive. .. The cells were grown in Dulbecco's modified Eagle medium (DMEM) (Sigma-Aldrich, St. Louis, MO) containing 0.75 mg/ml NaHCO3 , 20 mM HEPES (EuroClone, Milan, Italy), 1× PEST (100 IU/ml penicillin G, 100 μg/ml streptomycin sulfate; Gibco, Carlsbad, CA) and 5% fetal bovine serum (FBS) (Gibco) at 37°C.

Multiple Displacement Amplification:

Article Title: The High Mobility Group A1 (HMGA1) Chromatin Architectural Factor Modulates Nuclear Stiffness in Breast Cancer Cells
Article Snippet: Cells were cultured as previously described [ , ]: MDA-MB-231 and MDA-MB-157 cells were grown in DMEM supplemented with 2 mM L-glutamine, 10% tetracycline free fetal bovine serum (Tet-free FBS, Euroclone, Pero, Italy), penicillin (100 U/mL, Euroclone), streptomycin (100 μg/mL, Euroclone). .. MCF7 cells were grown in Dulbecco’s MEM Nutrient Mix F12 (1:1) with 25 mM HEPES (DME/F12-HEPES, Euroclone) supplemented with 2 mM L-glutamine, 10% tetracycline free fetal bovine serum (Tet-free FBS, Euroclone), penicillin (100 U/mL, Euroclone), streptomycin (100 μg/mL, Euroclone), and 1X MEM non-essential amino acids (Sigma Aldrich, St. Louis, MO, USA).

Isolation:

Article Title: Pro inflammatory stimuli enhance the immunosuppressive functions of adipose mesenchymal stem cells-derived exosomes
Article Snippet: Paragraph title: Monocytes isolation and differentiation into M1 macrophages ... Monocytes were separated from PBMCs by negative selection using a human CD14+ cell enrichment kit (StemCell Technologies) according to the manufacturer’s instructions and resuspended in RPMI medium supplemented with 10% heat inactivated fetal bovine serum (FBS), 1% glutamine, 1% pyruvate, 1% non-essential aminoacid, 1% penicillin/streptomycin, 1% Hepes (all from Euroclone).

Article Title: A Sialic Acid Binding Site in a Human Picornavirus
Article Snippet: Sample preparation The CVA24v strain (110390) used in this work originate from Malaysia and was isolated during the 2002–2004 pandemic . .. Briefly, normal human conjunctival (NHC) cells were infected with CVA24v in serumfree medium containing Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich), HEPES (pH 7.4; EuroClone, Milan, Italy), and penicillin-streptomycin (PEST; Gibco, Carlsbad, CA) with gentle agitation for one hour at 37°C.

Article Title: Immunosuppression by monocytic myeloid-derived suppressor cells in patients with pancreatic ductal carcinoma is orchestrated by STAT3
Article Snippet: Human proliferation assay PBMCs were isolated from leukocyte-enriched buffy coats from healthy volunteers (Transfusion Center, University and Hospital Trust of Verona, Verona, Italy) by Ficoll-Hypaque (GE Healthcare, Uppsala, Sweden) gradient centrifugation. .. Cell cultures were incubated at 37 °C and 8% CO2 in arginine and glutamine–Free-RPMI (Biochrom AG, Berlin, Germany), supplemented with 2 mM L-glutamine (Euroclone, Milano, Italy), 150 μM arginine (Sigma-Aldrich, St. Louis, MO, USA), 10% FBS (Superior, Merck, Darmstadt, Germany), 10 U/ml penicillin and streptomycin (Euroclone, Milano, Italy), and 0.1 mM HEPES (Euroclone, Milano, Italy).

Article Title: Induction of immunosuppressive functions and NF-κB by FLIP in monocytes
Article Snippet: Human proliferation assay PBMCs were isolated from leukocyte-enriched buffy coats from the healthy volunteer (Transfusion Center, University and Hospital Trust of Verona, Verona, Italy) by Ficoll-Hypaque (GE Healthcare, Uppsala, Sweden) gradient centrifugation as described above. .. PBMCs were recovered and washed in IMDM medium (Lonza, Visp, Switzerland), supplemented with 10% FBS (Euroclone, Milano, Italy), 100U/ml penicillin/streptomycin (Euroclone, Milano, Italy), β-mercaptoethanol (Sigma-Aldrich, Milan, Italy) and 10 mM HEPES (Euroclone, Milano, Italy).

Subcloning:

Article Title: The High Mobility Group A1 (HMGA1) Chromatin Architectural Factor Modulates Nuclear Stiffness in Breast Cancer Cells
Article Snippet: MCF7 cells were grown in Dulbecco’s MEM Nutrient Mix F12 (1:1) with 25 mM HEPES (DME/F12-HEPES, Euroclone) supplemented with 2 mM L-glutamine, 10% tetracycline free fetal bovine serum (Tet-free FBS, Euroclone), penicillin (100 U/mL, Euroclone), streptomycin (100 μg/mL, Euroclone), and 1X MEM non-essential amino acids (Sigma Aldrich, St. Louis, MO, USA). .. HMGA1 expression vector was obtained by subcloning the PCR products of the coding regions of human HMGA1a cDNA into the pcDNA3-HA plasmid expression vector (Invitrogen). pcDNA3-HA empty vector was used as a control.

Transfection:

Article Title: The High Mobility Group A1 (HMGA1) Chromatin Architectural Factor Modulates Nuclear Stiffness in Breast Cancer Cells
Article Snippet: MCF7 cells were grown in Dulbecco’s MEM Nutrient Mix F12 (1:1) with 25 mM HEPES (DME/F12-HEPES, Euroclone) supplemented with 2 mM L-glutamine, 10% tetracycline free fetal bovine serum (Tet-free FBS, Euroclone), penicillin (100 U/mL, Euroclone), streptomycin (100 μg/mL, Euroclone), and 1X MEM non-essential amino acids (Sigma Aldrich, St. Louis, MO, USA). .. Silencing experiments were carried out essentially as previously described [ ]; briefly, cell lines were transfected with 100 nM siRNAs with LipofectamineTM RNAiMAX reagent (Invitrogen, Carlsbad, CA, USA) and collected after 72 h. siRNA (Eurofins) sequences: HMGA1 (A1_3): ACTGGAGAAGGAGGAAGAG; CTRL: ACAGTCGCGTTTGCGACTG.

Labeling:

Article Title: Induction of immunosuppressive functions and NF-κB by FLIP in monocytes
Article Snippet: PBMCs were recovered and washed in IMDM medium (Lonza, Visp, Switzerland), supplemented with 10% FBS (Euroclone, Milano, Italy), 100U/ml penicillin/streptomycin (Euroclone, Milano, Italy), β-mercaptoethanol (Sigma-Aldrich, Milan, Italy) and 10 mM HEPES (Euroclone, Milano, Italy). .. Labeled “target” PBMCs were stimulated with coated 0.6 μg/ml anti-CD3 (clone OKT-3, eBioscience, Thermo Fisher Scientific, Waltham, MA, USA) and 5 μg/ml soluble anti-CD28 (clone CD28.2, eBioscience, Thermo Fisher Scientific, Waltham, MA, USA) for four days and co-cultured with “effectors” CD34+ or CD14+ cells at 0.1:1, 0.5:1, 1:1, 2:1, 3:1 ratio (effector: target) in 384 flat bottom well plates (BD, Franklin Lakes, NJ, USA).

Mouse Assay:

Article Title: Mast Cells Respond to Candida albicans Infections and Modulate Macrophages Phagocytosis of the Fungus
Article Snippet: BMMCs generation Bone marrow-derived mast cells (BMMCs) were obtained from 6- to 8- weeks-old mice by in vitro differentiation of bone marrow-derived progenitors obtained from mice femurs and tibiae. .. Precursor cells were cultivated in complete–IL-3 medium [RPMI 1,640 medium (Euroclone) supplemented with 20% FBS (Sigma Aldrich), 100 U·ml−1 penicillin, 100 μg·ml−1 Streptomycin, 2 mM Glutamine, 20 mM Hepes, non-essential amino acid, 1 mM Sodium Pyruvate (Euroclone), 50 mM β-mercaptoethanol (Sigma Aldrich), and 20 ng·ml−1 IL-3 (Peprotech)] at 37°C in 5% CO2 atmosphere.

Polymerase Chain Reaction:

Article Title: The High Mobility Group A1 (HMGA1) Chromatin Architectural Factor Modulates Nuclear Stiffness in Breast Cancer Cells
Article Snippet: MCF7 cells were grown in Dulbecco’s MEM Nutrient Mix F12 (1:1) with 25 mM HEPES (DME/F12-HEPES, Euroclone) supplemented with 2 mM L-glutamine, 10% tetracycline free fetal bovine serum (Tet-free FBS, Euroclone), penicillin (100 U/mL, Euroclone), streptomycin (100 μg/mL, Euroclone), and 1X MEM non-essential amino acids (Sigma Aldrich, St. Louis, MO, USA). .. HMGA1 expression vector was obtained by subcloning the PCR products of the coding regions of human HMGA1a cDNA into the pcDNA3-HA plasmid expression vector (Invitrogen). pcDNA3-HA empty vector was used as a control.

Plasmid Preparation:

Article Title: The High Mobility Group A1 (HMGA1) Chromatin Architectural Factor Modulates Nuclear Stiffness in Breast Cancer Cells
Article Snippet: MCF7 cells were grown in Dulbecco’s MEM Nutrient Mix F12 (1:1) with 25 mM HEPES (DME/F12-HEPES, Euroclone) supplemented with 2 mM L-glutamine, 10% tetracycline free fetal bovine serum (Tet-free FBS, Euroclone), penicillin (100 U/mL, Euroclone), streptomycin (100 μg/mL, Euroclone), and 1X MEM non-essential amino acids (Sigma Aldrich, St. Louis, MO, USA). .. HMGA1 expression vector was obtained by subcloning the PCR products of the coding regions of human HMGA1a cDNA into the pcDNA3-HA plasmid expression vector (Invitrogen). pcDNA3-HA empty vector was used as a control.

Functional Assay:

Article Title: Th17 Immunity in Children with Allergic Asthma and Rhinitis: A Pharmacological Approach
Article Snippet: Epithelial cell line culture 16-HBE (a differentiated SV-40 transformed bronchial epithelial cell) and human RPMI-2650 (squamous cell carcinoma of the nasal septum) (American Type Culture Collection, ATCC, Manassas, VA, USA) cell lines represent a valid in vitro model to evaluate the functional properties of bronchial and nasal epithelial cells in inflammation and repair processes , . .. Cells were maintained in MEM (Gibco) supplemented with 10% foetal bovine serum (FBS) (Hyclone), 2mM glutamine (Euroclone), Hepes (Euroclone), gentamicin 250 μg/ml (Gibco) and 1% MEM non essential aminoacids (Gibco).

Selection:

Article Title: Pro inflammatory stimuli enhance the immunosuppressive functions of adipose mesenchymal stem cells-derived exosomes
Article Snippet: .. Monocytes were separated from PBMCs by negative selection using a human CD14+ cell enrichment kit (StemCell Technologies) according to the manufacturer’s instructions and resuspended in RPMI medium supplemented with 10% heat inactivated fetal bovine serum (FBS), 1% glutamine, 1% pyruvate, 1% non-essential aminoacid, 1% penicillin/streptomycin, 1% Hepes (all from Euroclone). ..

Sample Prep:

Article Title: A Sialic Acid Binding Site in a Human Picornavirus
Article Snippet: Paragraph title: Sample preparation ... Briefly, normal human conjunctival (NHC) cells were infected with CVA24v in serumfree medium containing Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich), HEPES (pH 7.4; EuroClone, Milan, Italy), and penicillin-streptomycin (PEST; Gibco, Carlsbad, CA) with gentle agitation for one hour at 37°C.

In Vitro:

Article Title: Facile Preparation of N-Glycosylated 10-Piperazinyl Artemisinin Derivatives and Evaluation of Their Antimalarial and Cytotoxic Activities
Article Snippet: Paragraph title: 4.4.1. Cultures of Pf and In Vitro Antimalarial Assays ... Parasites were maintained at 5% hematocrit (human type A-positive red blood cells) in RPMI 1640 (EuroClone, Celbio) medium with the addition of 10% heat inactivate human serum, 20 mM Hepes and 2 mM glutamine (Euroclone).

Article Title: Th17 Immunity in Children with Allergic Asthma and Rhinitis: A Pharmacological Approach
Article Snippet: Epithelial cell line culture 16-HBE (a differentiated SV-40 transformed bronchial epithelial cell) and human RPMI-2650 (squamous cell carcinoma of the nasal septum) (American Type Culture Collection, ATCC, Manassas, VA, USA) cell lines represent a valid in vitro model to evaluate the functional properties of bronchial and nasal epithelial cells in inflammation and repair processes , . .. Cells were maintained in MEM (Gibco) supplemented with 10% foetal bovine serum (FBS) (Hyclone), 2mM glutamine (Euroclone), Hepes (Euroclone), gentamicin 250 μg/ml (Gibco) and 1% MEM non essential aminoacids (Gibco).

Article Title: Mast Cells Respond to Candida albicans Infections and Modulate Macrophages Phagocytosis of the Fungus
Article Snippet: BMMCs generation Bone marrow-derived mast cells (BMMCs) were obtained from 6- to 8- weeks-old mice by in vitro differentiation of bone marrow-derived progenitors obtained from mice femurs and tibiae. .. Precursor cells were cultivated in complete–IL-3 medium [RPMI 1,640 medium (Euroclone) supplemented with 20% FBS (Sigma Aldrich), 100 U·ml−1 penicillin, 100 μg·ml−1 Streptomycin, 2 mM Glutamine, 20 mM Hepes, non-essential amino acid, 1 mM Sodium Pyruvate (Euroclone), 50 mM β-mercaptoethanol (Sigma Aldrich), and 20 ng·ml−1 IL-3 (Peprotech)] at 37°C in 5% CO2 atmosphere.

Concentration Assay:

Article Title: Facile Preparation of N-Glycosylated 10-Piperazinyl Artemisinin Derivatives and Evaluation of Their Antimalarial and Cytotoxic Activities
Article Snippet: Parasites were maintained at 5% hematocrit (human type A-positive red blood cells) in RPMI 1640 (EuroClone, Celbio) medium with the addition of 10% heat inactivate human serum, 20 mM Hepes and 2 mM glutamine (Euroclone). .. Compounds were dissolved in either water or DMSO and then diluted with medium to achieve the required concentrations (final DMSO concentration < 1%, which is non-toxic to the parasite).

Article Title: A Sialic Acid Binding Site in a Human Picornavirus
Article Snippet: Briefly, normal human conjunctival (NHC) cells were infected with CVA24v in serumfree medium containing Dulbecco's modified Eagle's medium (DMEM; Sigma-Aldrich), HEPES (pH 7.4; EuroClone, Milan, Italy), and penicillin-streptomycin (PEST; Gibco, Carlsbad, CA) with gentle agitation for one hour at 37°C. .. Thirty hours after infection, Triton X-100 (Sigma-Aldrich) was added to a final concentration of 0.5% and centrifuged for 15 min at 3,000×g .

Article Title: Immunosuppression by monocytic myeloid-derived suppressor cells in patients with pancreatic ductal carcinoma is orchestrated by STAT3
Article Snippet: PBMCs were recovered, washed in IMDM medium (Lonza, Visp, Switzerland), supplemented with 10% FBS (Euroclone, Milano, Italy), 100 U/ml penicillin/streptomycin (Euroclone, Milano, Italy), β-mercaptoethanol (Sigma-Aldrich, Milan, Italy) and 10 mM HEPES (Euroclone, Milano, Italy), resuspended at a final concentration of 107 cells/ml in PBS and stained with 1 μM as final working concentration of CellTrace Violet stock solution (Thermo Fisher Scientific, Waltham, MA, USA), followed by 5 min’ incubation at 37 °C, protected from light. .. Cell cultures were incubated at 37 °C and 8% CO2 in arginine and glutamine–Free-RPMI (Biochrom AG, Berlin, Germany), supplemented with 2 mM L-glutamine (Euroclone, Milano, Italy), 150 μM arginine (Sigma-Aldrich, St. Louis, MO, USA), 10% FBS (Superior, Merck, Darmstadt, Germany), 10 U/ml penicillin and streptomycin (Euroclone, Milano, Italy), and 0.1 mM HEPES (Euroclone, Milano, Italy).

Article Title: Induction of immunosuppressive functions and NF-κB by FLIP in monocytes
Article Snippet: PBMCs were recovered and washed in IMDM medium (Lonza, Visp, Switzerland), supplemented with 10% FBS (Euroclone, Milano, Italy), 100U/ml penicillin/streptomycin (Euroclone, Milano, Italy), β-mercaptoethanol (Sigma-Aldrich, Milan, Italy) and 10 mM HEPES (Euroclone, Milano, Italy). .. PBMCs were resuspended at a final concentration of 107 cells/ml in PBS and stained with 2,5 μM as final working concentration of CellTrace Violet stock solution (Thermo Fisher Scientific, Waltham, MA, USA), followed by 5 min’ incubation at 37 °C, protected from light.

Staining:

Article Title: Pro inflammatory stimuli enhance the immunosuppressive functions of adipose mesenchymal stem cells-derived exosomes
Article Snippet: Monocytes were separated from PBMCs by negative selection using a human CD14+ cell enrichment kit (StemCell Technologies) according to the manufacturer’s instructions and resuspended in RPMI medium supplemented with 10% heat inactivated fetal bovine serum (FBS), 1% glutamine, 1% pyruvate, 1% non-essential aminoacid, 1% penicillin/streptomycin, 1% Hepes (all from Euroclone). .. Purity of monocytes was over 95% as judged by staining with anti-CD14 (eBiosciences) (data not shown).

Article Title: Immunosuppression by monocytic myeloid-derived suppressor cells in patients with pancreatic ductal carcinoma is orchestrated by STAT3
Article Snippet: PBMCs were recovered, washed in IMDM medium (Lonza, Visp, Switzerland), supplemented with 10% FBS (Euroclone, Milano, Italy), 100 U/ml penicillin/streptomycin (Euroclone, Milano, Italy), β-mercaptoethanol (Sigma-Aldrich, Milan, Italy) and 10 mM HEPES (Euroclone, Milano, Italy), resuspended at a final concentration of 107 cells/ml in PBS and stained with 1 μM as final working concentration of CellTrace Violet stock solution (Thermo Fisher Scientific, Waltham, MA, USA), followed by 5 min’ incubation at 37 °C, protected from light. .. Cell cultures were incubated at 37 °C and 8% CO2 in arginine and glutamine–Free-RPMI (Biochrom AG, Berlin, Germany), supplemented with 2 mM L-glutamine (Euroclone, Milano, Italy), 150 μM arginine (Sigma-Aldrich, St. Louis, MO, USA), 10% FBS (Superior, Merck, Darmstadt, Germany), 10 U/ml penicillin and streptomycin (Euroclone, Milano, Italy), and 0.1 mM HEPES (Euroclone, Milano, Italy).

Article Title: Mast Cells Respond to Candida albicans Infections and Modulate Macrophages Phagocytosis of the Fungus
Article Snippet: Precursor cells were cultivated in complete–IL-3 medium [RPMI 1,640 medium (Euroclone) supplemented with 20% FBS (Sigma Aldrich), 100 U·ml−1 penicillin, 100 μg·ml−1 Streptomycin, 2 mM Glutamine, 20 mM Hepes, non-essential amino acid, 1 mM Sodium Pyruvate (Euroclone), 50 mM β-mercaptoethanol (Sigma Aldrich), and 20 ng·ml−1 IL-3 (Peprotech)] at 37°C in 5% CO2 atmosphere. .. After 5 weeks, BMMCs differentiation was confirmed by flow cytometry by staining with fluorochrome-conjugated anti-FcεRIa (MAR-1) and anti-cKit (ACK2) antibodies (eBiosciences).

Article Title: Induction of immunosuppressive functions and NF-κB by FLIP in monocytes
Article Snippet: PBMCs were recovered and washed in IMDM medium (Lonza, Visp, Switzerland), supplemented with 10% FBS (Euroclone, Milano, Italy), 100U/ml penicillin/streptomycin (Euroclone, Milano, Italy), β-mercaptoethanol (Sigma-Aldrich, Milan, Italy) and 10 mM HEPES (Euroclone, Milano, Italy). .. PBMCs were resuspended at a final concentration of 107 cells/ml in PBS and stained with 2,5 μM as final working concentration of CellTrace Violet stock solution (Thermo Fisher Scientific, Waltham, MA, USA), followed by 5 min’ incubation at 37 °C, protected from light.

Gradient Centrifugation:

Article Title: Pro inflammatory stimuli enhance the immunosuppressive functions of adipose mesenchymal stem cells-derived exosomes
Article Snippet: Human peripheral blood mononuclear cells (PBMCs) were isolated from EDTA-uncoagulated blood of blood donors by Ficoll gradient centrifugation (Millipore). .. Monocytes were separated from PBMCs by negative selection using a human CD14+ cell enrichment kit (StemCell Technologies) according to the manufacturer’s instructions and resuspended in RPMI medium supplemented with 10% heat inactivated fetal bovine serum (FBS), 1% glutamine, 1% pyruvate, 1% non-essential aminoacid, 1% penicillin/streptomycin, 1% Hepes (all from Euroclone).

Article Title: Immunosuppression by monocytic myeloid-derived suppressor cells in patients with pancreatic ductal carcinoma is orchestrated by STAT3
Article Snippet: Human proliferation assay PBMCs were isolated from leukocyte-enriched buffy coats from healthy volunteers (Transfusion Center, University and Hospital Trust of Verona, Verona, Italy) by Ficoll-Hypaque (GE Healthcare, Uppsala, Sweden) gradient centrifugation. .. Cell cultures were incubated at 37 °C and 8% CO2 in arginine and glutamine–Free-RPMI (Biochrom AG, Berlin, Germany), supplemented with 2 mM L-glutamine (Euroclone, Milano, Italy), 150 μM arginine (Sigma-Aldrich, St. Louis, MO, USA), 10% FBS (Superior, Merck, Darmstadt, Germany), 10 U/ml penicillin and streptomycin (Euroclone, Milano, Italy), and 0.1 mM HEPES (Euroclone, Milano, Italy).

Article Title: Induction of immunosuppressive functions and NF-κB by FLIP in monocytes
Article Snippet: Human proliferation assay PBMCs were isolated from leukocyte-enriched buffy coats from the healthy volunteer (Transfusion Center, University and Hospital Trust of Verona, Verona, Italy) by Ficoll-Hypaque (GE Healthcare, Uppsala, Sweden) gradient centrifugation as described above. .. PBMCs were recovered and washed in IMDM medium (Lonza, Visp, Switzerland), supplemented with 10% FBS (Euroclone, Milano, Italy), 100U/ml penicillin/streptomycin (Euroclone, Milano, Italy), β-mercaptoethanol (Sigma-Aldrich, Milan, Italy) and 10 mM HEPES (Euroclone, Milano, Italy).

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    EuroClone hepes
    Hepes, supplied by EuroClone, used in various techniques. Bioz Stars score: 94/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    EuroClone hepes buffer
    Hepes Buffer, supplied by EuroClone, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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