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Millipore t25 flask
T25 Flask, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 6 article reviews
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t25 flask - by Bioz Stars, 2020-04
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Methylation Sequencing:

Article Title: Methylation of a Single Intronic CpG Mediates Expression Silencing of the PMP24 Gene in Prostate Cancer
Article Snippet: For the demethylation assay, PC-3 cells were seeded at a density of 1 × 105 cells per T25 flask at day 0, one day before treatment with 5-aza-dC (Sigma, St. Louis, MO). .. The cells were collected at day 6 for bisulfite sequencing and real-time RT-PCR analyses.

Clone Assay:

Article Title: Fanconi Anemia Gene Editing by the CRISPR/Cas9 System
Article Snippet: PCR products were cloned into the pCR 4 TOPO vector (Invitrogen) for Sanger sequencing confirmation of the entire amplicon using the M13 forward and reverse primers. .. H2AX staining was performed on cells seeded at a concentration of 120,000 total cells in a T25 flask in the presence of 2 m M hydroxyurea (Sigma) for 48 hr using the H2AX phosphorylation assay kit according to the manufacturer's instructions (EMD Millipore, Billerica, MA).

Centrifugation:

Article Title: Palmitoylation of Desmoglein 2 Is a Regulator of Assembly Dynamics and Protein Turnover *
Article Snippet: Cells grown in a T25 flask were scraped into 1 ml of Triton X-100-containing buffer (10 m m Tris-HCl (pH 7.0), 0.5% Triton X-100 (Sigma), 5 m m EDTA, 2 m m EGTA, 30 m m sodium fluoride, 40 m m β-glycerophosphate, 10 m m sodium pyrophosphate, 2 m m sodium orthovanadate, and 2 m m phenylmethylsulfonyl fluoride) and incubated on ice while shaking for 15 min. .. The insoluble material was collected by centrifugation for 15 min at 14,000 × g .

Amplification:

Article Title: Fanconi Anemia Gene Editing by the CRISPR/Cas9 System
Article Snippet: PCR products were cloned into the pCR 4 TOPO vector (Invitrogen) for Sanger sequencing confirmation of the entire amplicon using the M13 forward and reverse primers. .. H2AX staining was performed on cells seeded at a concentration of 120,000 total cells in a T25 flask in the presence of 2 m M hydroxyurea (Sigma) for 48 hr using the H2AX phosphorylation assay kit according to the manufacturer's instructions (EMD Millipore, Billerica, MA).

Transduction:

Article Title: An Influenza Virus M2 Protein Specific Chimeric Antigen Receptor Modulates Influenza A/WSN/33 H1N1 Infection In Vivo
Article Snippet: Paragraph title: Transduction of Murine Splenocytes ... Splenocytes were seeded at 5x106 /ml in a T25 flask before addition of 2.5 μg/ml Concanavalin A (Calbiochem), 1ng/ml IL-7 (Santa Cruz) and 100 U/ml human rIL-2, and incubated for 24 hours.

Cytometry:

Article Title: Fanconi Anemia Gene Editing by the CRISPR/Cas9 System
Article Snippet: H2AX staining was performed on cells seeded at a concentration of 120,000 total cells in a T25 flask in the presence of 2 m M hydroxyurea (Sigma) for 48 hr using the H2AX phosphorylation assay kit according to the manufacturer's instructions (EMD Millipore, Billerica, MA). .. Flow cytometry was performed using the BD LSRFortessa Cell Analyzer (BD Biosciences).

Article Title: An Influenza Virus M2 Protein Specific Chimeric Antigen Receptor Modulates Influenza A/WSN/33 H1N1 Infection In Vivo
Article Snippet: Splenocytes were seeded at 5x106 /ml in a T25 flask before addition of 2.5 μg/ml Concanavalin A (Calbiochem), 1ng/ml IL-7 (Santa Cruz) and 100 U/ml human rIL-2, and incubated for 24 hours. .. The transduced splenocytes were cultured in fresh culture media with 100 U/ml IL-2, at a cell density of 0.15×106 /ml in 2 ml per well of a 24 well plate for 2 days before being assessed for transduction efficiency using flow cytometry and a CD34 antibody.

Quantitative RT-PCR:

Article Title: Methylation of a Single Intronic CpG Mediates Expression Silencing of the PMP24 Gene in Prostate Cancer
Article Snippet: For the demethylation assay, PC-3 cells were seeded at a density of 1 × 105 cells per T25 flask at day 0, one day before treatment with 5-aza-dC (Sigma, St. Louis, MO). .. The cells were collected at day 6 for bisulfite sequencing and real-time RT-PCR analyses.

Real-time Polymerase Chain Reaction:

Article Title: Molecular damage and responses of oral keratinocyte to hydrogen peroxide
Article Snippet: The tube was centrifuged again at 1200 rpm for 5 min and the pellet was resuspended with KGM (5 mL) and then incubated in a T25 flask containing 100X lincomycin (10 mg/mL) (Sigma-Aldrich, St. Louis, MO, USA). .. The NHOKs were characterized periodically for microscopic presentation and their intermediate filament expression profile (keratin+, vimentin-) as well as free of mycoplasma contamination by qPCR.

Incubation:

Article Title: An Influenza Virus M2 Protein Specific Chimeric Antigen Receptor Modulates Influenza A/WSN/33 H1N1 Infection In Vivo
Article Snippet: .. Splenocytes were seeded at 5x106 /ml in a T25 flask before addition of 2.5 μg/ml Concanavalin A (Calbiochem), 1ng/ml IL-7 (Santa Cruz) and 100 U/ml human rIL-2, and incubated for 24 hours. .. The activated splenocytes were then transduced with retrovirus harvested from the packaging cells using retronectin coated plates as described above.

Article Title: Effect of Hyaluronic Acid Hydrogels Containing Astrocyte-Derived Extracellular Matrix and/or V2a Interneurons on Histologic Outcomes following Spinal Cord Injury
Article Snippet: 0.25% Trypsin-EDTA (Life Technologies) incubation at 37°C for 5 min was used to dissociate mESCs from the culture flask. .. This reaction was then quenched with fresh CM and cells were seeded into a new T25 flask coated with 0.1% gelatin (Sigma).

Article Title: Mycoplasma gallisepticum Lipid Associated Membrane Proteins Up-regulate Inflammatory Genes in Chicken Tracheal Epithelial Cells via TLR-2 Ligation through an NF-κB Dependent Pathway
Article Snippet: To dissociate the epithelial cells from the underlying connective tissue, about 3–4 tracheal pieces were placed in a T25 flask containing 25 ml dissociation solution [DMEM 50 ml, protease type XIV 0.14 g (Sigma Aldrich), DNAse 0.01 g (Sigma Aldrich) and antibiotic/antimycotic 1X (Gibco, Life technologies)]. .. 10% FBS (Gibco, Life technologies) was added to block protease activity after incubation.

Article Title: Molecular damage and responses of oral keratinocyte to hydrogen peroxide
Article Snippet: .. The tube was centrifuged again at 1200 rpm for 5 min and the pellet was resuspended with KGM (5 mL) and then incubated in a T25 flask containing 100X lincomycin (10 mg/mL) (Sigma-Aldrich, St. Louis, MO, USA). ..

Article Title: Palmitoylation of Desmoglein 2 Is a Regulator of Assembly Dynamics and Protein Turnover *
Article Snippet: .. Cells grown in a T25 flask were scraped into 1 ml of Triton X-100-containing buffer (10 m m Tris-HCl (pH 7.0), 0.5% Triton X-100 (Sigma), 5 m m EDTA, 2 m m EGTA, 30 m m sodium fluoride, 40 m m β-glycerophosphate, 10 m m sodium pyrophosphate, 2 m m sodium orthovanadate, and 2 m m phenylmethylsulfonyl fluoride) and incubated on ice while shaking for 15 min. .. The insoluble material was collected by centrifugation for 15 min at 14,000 × g .

Activity Assay:

Article Title: Mycoplasma gallisepticum Lipid Associated Membrane Proteins Up-regulate Inflammatory Genes in Chicken Tracheal Epithelial Cells via TLR-2 Ligation through an NF-κB Dependent Pathway
Article Snippet: To dissociate the epithelial cells from the underlying connective tissue, about 3–4 tracheal pieces were placed in a T25 flask containing 25 ml dissociation solution [DMEM 50 ml, protease type XIV 0.14 g (Sigma Aldrich), DNAse 0.01 g (Sigma Aldrich) and antibiotic/antimycotic 1X (Gibco, Life technologies)]. .. 10% FBS (Gibco, Life technologies) was added to block protease activity after incubation.

Expressing:

Article Title: Molecular damage and responses of oral keratinocyte to hydrogen peroxide
Article Snippet: The tube was centrifuged again at 1200 rpm for 5 min and the pellet was resuspended with KGM (5 mL) and then incubated in a T25 flask containing 100X lincomycin (10 mg/mL) (Sigma-Aldrich, St. Louis, MO, USA). .. The NHOKs were characterized periodically for microscopic presentation and their intermediate filament expression profile (keratin+, vimentin-) as well as free of mycoplasma contamination by qPCR.

Article Title: Transgenic Enrichment of Mouse Embryonic Stem Cell-derived Progenitor Motor Neurons
Article Snippet: Olig2 expression was characterized using the G-Olig2 ESC line that expresses GFP under the control of the Olig2 GRE [ - ]. .. Cells were passaged at a 1:5 ratio every 2 days and seeded on a new T25 flask coated with a 0.1% gelatin solution (Sigma).

Modification:

Article Title: Mycoplasma gallisepticum Lipid Associated Membrane Proteins Up-regulate Inflammatory Genes in Chicken Tracheal Epithelial Cells via TLR-2 Ligation through an NF-κB Dependent Pathway
Article Snippet: The tracheae were rinsed twice in DMEM (Dulbecco's modified eagles medium) (Gibco, Life technologies; Grand Island, NY) under sterile conditions. .. To dissociate the epithelial cells from the underlying connective tissue, about 3–4 tracheal pieces were placed in a T25 flask containing 25 ml dissociation solution [DMEM 50 ml, protease type XIV 0.14 g (Sigma Aldrich), DNAse 0.01 g (Sigma Aldrich) and antibiotic/antimycotic 1X (Gibco, Life technologies)].

Article Title: Transgenic Enrichment of Mouse Embryonic Stem Cell-derived Progenitor Motor Neurons
Article Snippet: ESCs were grown in complete media consisting of Dulbecco’s modified Eagle’s Medium (DMEM; Invitrogen, Carlsbad, CA, ) supplemented with 10% newborn calf serum (Invitrogen), 10% fetal bovine serum (Invitrogen), 10 μM thymidine (Sigma, St. Louis, MO, ), and 30 μM of each of the following nucleosides: adenosine, cytosine, guanosine, and uridine (Sigma). .. Cells were passaged at a 1:5 ratio every 2 days and seeded on a new T25 flask coated with a 0.1% gelatin solution (Sigma).

Derivative Assay:

Article Title: C9orf72 hypermethylation protects against repeat expansion-associated pathology in ALS/FTD
Article Snippet: Lymphoblast cell lines derived from either a repeat expansion carrier (ND14442) or a non-expanded ALS patient (ND16183) were grown in RPMI 1640 with 15 % fetal bovine serum and 2 mM L-glutamine in 37 °C with 5 % CO2 . .. Cells were seeded at a concentration of 300,000 cells/ml in 10 ml in a T25 flask, and 0.5 μM 5-aza-dC (Sigma-Aldrich, St. Louis, MO, USA) was spiked daily into lymphoblast cells for 3 days followed by a 3-day recovery period without 5-aza-dC.

Flow Cytometry:

Article Title: Fanconi Anemia Gene Editing by the CRISPR/Cas9 System
Article Snippet: H2AX staining was performed on cells seeded at a concentration of 120,000 total cells in a T25 flask in the presence of 2 m M hydroxyurea (Sigma) for 48 hr using the H2AX phosphorylation assay kit according to the manufacturer's instructions (EMD Millipore, Billerica, MA). .. Flow cytometry was performed using the BD LSRFortessa Cell Analyzer (BD Biosciences).

Article Title: An Influenza Virus M2 Protein Specific Chimeric Antigen Receptor Modulates Influenza A/WSN/33 H1N1 Infection In Vivo
Article Snippet: Splenocytes were seeded at 5x106 /ml in a T25 flask before addition of 2.5 μg/ml Concanavalin A (Calbiochem), 1ng/ml IL-7 (Santa Cruz) and 100 U/ml human rIL-2, and incubated for 24 hours. .. The transduced splenocytes were cultured in fresh culture media with 100 U/ml IL-2, at a cell density of 0.15×106 /ml in 2 ml per well of a 24 well plate for 2 days before being assessed for transduction efficiency using flow cytometry and a CD34 antibody.

Cell Culture:

Article Title: Multi-Electrode Array with a Planar Surface for Cell Patterning by Microprinting
Article Snippet: .. HL-1 cells were cultured on the plastic bottom of a T25 flask in Claycomb Medium, supplemented with 10% fetal bovine serum, 100 U/mL penicillin, 100 mg/mL streptomycin, 0.3 mM ascorbic acid, 10 mM norepinephrine, and 2 mM L-glutamine (all ingredients from Sigma-Aldrich), at 37 °C in a humid atmosphere of 5% CO2 . ..

Article Title: Methylation of a Single Intronic CpG Mediates Expression Silencing of the PMP24 Gene in Prostate Cancer
Article Snippet: Paragraph title: Cell Culture and 5-aza-dC Treatment ... For the demethylation assay, PC-3 cells were seeded at a density of 1 × 105 cells per T25 flask at day 0, one day before treatment with 5-aza-dC (Sigma, St. Louis, MO).

Article Title: An Influenza Virus M2 Protein Specific Chimeric Antigen Receptor Modulates Influenza A/WSN/33 H1N1 Infection In Vivo
Article Snippet: Splenocytes were seeded at 5x106 /ml in a T25 flask before addition of 2.5 μg/ml Concanavalin A (Calbiochem), 1ng/ml IL-7 (Santa Cruz) and 100 U/ml human rIL-2, and incubated for 24 hours. .. The transduced splenocytes were cultured in fresh culture media with 100 U/ml IL-2, at a cell density of 0.15×106 /ml in 2 ml per well of a 24 well plate for 2 days before being assessed for transduction efficiency using flow cytometry and a CD34 antibody.

Article Title: Mycoplasma gallisepticum Lipid Associated Membrane Proteins Up-regulate Inflammatory Genes in Chicken Tracheal Epithelial Cells via TLR-2 Ligation through an NF-κB Dependent Pathway
Article Snippet: Paragraph title: Cell culture and exposure/stimulation ... To dissociate the epithelial cells from the underlying connective tissue, about 3–4 tracheal pieces were placed in a T25 flask containing 25 ml dissociation solution [DMEM 50 ml, protease type XIV 0.14 g (Sigma Aldrich), DNAse 0.01 g (Sigma Aldrich) and antibiotic/antimycotic 1X (Gibco, Life technologies)].

Reverse Transcription Polymerase Chain Reaction:

Article Title: Fanconi Anemia Gene Editing by the CRISPR/Cas9 System
Article Snippet: For RT-PCR, total cellular RNA was isolated and reverse transcribed using SuperScript Vilo (Invitrogen) followed by amplification with FANCC allele-specific RT forward (5′-GGTGTATTAAGCCATATTCTGAGC-3′) and reverse (5′-ACAACCCGGAATATGGCAGG-3′). .. H2AX staining was performed on cells seeded at a concentration of 120,000 total cells in a T25 flask in the presence of 2 m M hydroxyurea (Sigma) for 48 hr using the H2AX phosphorylation assay kit according to the manufacturer's instructions (EMD Millipore, Billerica, MA).

Lactate Dehydrogenase Assay:

Article Title: C9orf72 hypermethylation protects against repeat expansion-associated pathology in ALS/FTD
Article Snippet: Cells were seeded at a concentration of 300,000 cells/ml in 10 ml in a T25 flask, and 0.5 μM 5-aza-dC (Sigma-Aldrich, St. Louis, MO, USA) was spiked daily into lymphoblast cells for 3 days followed by a 3-day recovery period without 5-aza-dC. .. After 24 h, cells and media were collected and measured in triplicate using a lactate dehydrogenase assay (Clontech, Mountain View, CA, USA) to calculate the percent LDH release into the media relative to total LDH from untreated cell pellets.

Generated:

Article Title: Methylation of a Single Intronic CpG Mediates Expression Silencing of the PMP24 Gene in Prostate Cancer
Article Snippet: An immortalized human normal prostate epithelial cell line (NPrEC) was generated in our laboratory and maintained as previously described [ ]. .. For the demethylation assay, PC-3 cells were seeded at a density of 1 × 105 cells per T25 flask at day 0, one day before treatment with 5-aza-dC (Sigma, St. Louis, MO).

Sequencing:

Article Title: Fanconi Anemia Gene Editing by the CRISPR/Cas9 System
Article Snippet: PCR products were cloned into the pCR 4 TOPO vector (Invitrogen) for Sanger sequencing confirmation of the entire amplicon using the M13 forward and reverse primers. .. H2AX staining was performed on cells seeded at a concentration of 120,000 total cells in a T25 flask in the presence of 2 m M hydroxyurea (Sigma) for 48 hr using the H2AX phosphorylation assay kit according to the manufacturer's instructions (EMD Millipore, Billerica, MA).

Isolation:

Article Title: Fanconi Anemia Gene Editing by the CRISPR/Cas9 System
Article Snippet: For RT-PCR, total cellular RNA was isolated and reverse transcribed using SuperScript Vilo (Invitrogen) followed by amplification with FANCC allele-specific RT forward (5′-GGTGTATTAAGCCATATTCTGAGC-3′) and reverse (5′-ACAACCCGGAATATGGCAGG-3′). .. H2AX staining was performed on cells seeded at a concentration of 120,000 total cells in a T25 flask in the presence of 2 m M hydroxyurea (Sigma) for 48 hr using the H2AX phosphorylation assay kit according to the manufacturer's instructions (EMD Millipore, Billerica, MA).

Article Title: An Influenza Virus M2 Protein Specific Chimeric Antigen Receptor Modulates Influenza A/WSN/33 H1N1 Infection In Vivo
Article Snippet: Spleens were isolated from Balb/c mice and passed through a 70 μm cell strainer into a 50 ml tube using 20mls of PBS. .. Splenocytes were seeded at 5x106 /ml in a T25 flask before addition of 2.5 μg/ml Concanavalin A (Calbiochem), 1ng/ml IL-7 (Santa Cruz) and 100 U/ml human rIL-2, and incubated for 24 hours.

Article Title: Molecular damage and responses of oral keratinocyte to hydrogen peroxide
Article Snippet: Gingival tissue was isolated from excess tissue during the surgical removal of impacted wisdom teeth from 10 healthy patients (age 19~50) under approval by NCKUH Institutional Review Board for Human Studies. .. The tube was centrifuged again at 1200 rpm for 5 min and the pellet was resuspended with KGM (5 mL) and then incubated in a T25 flask containing 100X lincomycin (10 mg/mL) (Sigma-Aldrich, St. Louis, MO, USA).

Article Title: Palmitoylation of Desmoglein 2 Is a Regulator of Assembly Dynamics and Protein Turnover *
Article Snippet: Triton X-100 soluble (s) and insoluble (p) fractions were isolated by first washing cell monolayers with 1× phosphate-buffered saline. .. Cells grown in a T25 flask were scraped into 1 ml of Triton X-100-containing buffer (10 m m Tris-HCl (pH 7.0), 0.5% Triton X-100 (Sigma), 5 m m EDTA, 2 m m EGTA, 30 m m sodium fluoride, 40 m m β-glycerophosphate, 10 m m sodium pyrophosphate, 2 m m sodium orthovanadate, and 2 m m phenylmethylsulfonyl fluoride) and incubated on ice while shaking for 15 min.

Protein Quantitation:

Article Title: De novo lipogenesis in the differentiating human adipocyte can provide all fatty acids necessary for maturation [S]
Article Snippet: Paragraph title: Cellular protein quantitation ... Total adipocyte protein per T25 flask was determined with the Bradford method ( ) using Coomassie Blue G (Sigma-Aldrich, Gillingham, UK) made up according to the manufacturer's instructions.

Transfection:

Article Title: An Influenza Virus M2 Protein Specific Chimeric Antigen Receptor Modulates Influenza A/WSN/33 H1N1 Infection In Vivo
Article Snippet: Transduction of Murine Splenocytes The packaging cells (293T/Phoenix Amp) were prepared and transfected as above except that the pColtGalV plasmid was replaced with pHIT123 (MLV-E Env, provided by Prof. R. Debets, University of Rotterdam). .. Splenocytes were seeded at 5x106 /ml in a T25 flask before addition of 2.5 μg/ml Concanavalin A (Calbiochem), 1ng/ml IL-7 (Santa Cruz) and 100 U/ml human rIL-2, and incubated for 24 hours.

Mouse Assay:

Article Title: Multi-Electrode Array with a Planar Surface for Cell Patterning by Microprinting
Article Snippet: HL-1 Cell Culturing, Measurements of Action Potentials, Staining, and Visualization HL-1 cells (cardiac myocytes from transgenic mice) were obtained from Sigma Aldrich. .. HL-1 cells were cultured on the plastic bottom of a T25 flask in Claycomb Medium, supplemented with 10% fetal bovine serum, 100 U/mL penicillin, 100 mg/mL streptomycin, 0.3 mM ascorbic acid, 10 mM norepinephrine, and 2 mM L-glutamine (all ingredients from Sigma-Aldrich), at 37 °C in a humid atmosphere of 5% CO2 .

Article Title: An Influenza Virus M2 Protein Specific Chimeric Antigen Receptor Modulates Influenza A/WSN/33 H1N1 Infection In Vivo
Article Snippet: Spleens were isolated from Balb/c mice and passed through a 70 μm cell strainer into a 50 ml tube using 20mls of PBS. .. Splenocytes were seeded at 5x106 /ml in a T25 flask before addition of 2.5 μg/ml Concanavalin A (Calbiochem), 1ng/ml IL-7 (Santa Cruz) and 100 U/ml human rIL-2, and incubated for 24 hours.

Polymerase Chain Reaction:

Article Title: Fanconi Anemia Gene Editing by the CRISPR/Cas9 System
Article Snippet: PCR products were cloned into the pCR 4 TOPO vector (Invitrogen) for Sanger sequencing confirmation of the entire amplicon using the M13 forward and reverse primers. .. H2AX staining was performed on cells seeded at a concentration of 120,000 total cells in a T25 flask in the presence of 2 m M hydroxyurea (Sigma) for 48 hr using the H2AX phosphorylation assay kit according to the manufacturer's instructions (EMD Millipore, Billerica, MA).

Blocking Assay:

Article Title: Mycoplasma gallisepticum Lipid Associated Membrane Proteins Up-regulate Inflammatory Genes in Chicken Tracheal Epithelial Cells via TLR-2 Ligation through an NF-κB Dependent Pathway
Article Snippet: To dissociate the epithelial cells from the underlying connective tissue, about 3–4 tracheal pieces were placed in a T25 flask containing 25 ml dissociation solution [DMEM 50 ml, protease type XIV 0.14 g (Sigma Aldrich), DNAse 0.01 g (Sigma Aldrich) and antibiotic/antimycotic 1X (Gibco, Life technologies)]. .. 10% FBS (Gibco, Life technologies) was added to block protease activity after incubation.

Staining:

Article Title: Fanconi Anemia Gene Editing by the CRISPR/Cas9 System
Article Snippet: .. H2AX staining was performed on cells seeded at a concentration of 120,000 total cells in a T25 flask in the presence of 2 m M hydroxyurea (Sigma) for 48 hr using the H2AX phosphorylation assay kit according to the manufacturer's instructions (EMD Millipore, Billerica, MA). .. Flow cytometry was performed using the BD LSRFortessa Cell Analyzer (BD Biosciences).

Article Title: Multi-Electrode Array with a Planar Surface for Cell Patterning by Microprinting
Article Snippet: Paragraph title: 2.4. HL-1 Cell Culturing, Measurements of Action Potentials, Staining, and Visualization ... HL-1 cells were cultured on the plastic bottom of a T25 flask in Claycomb Medium, supplemented with 10% fetal bovine serum, 100 U/mL penicillin, 100 mg/mL streptomycin, 0.3 mM ascorbic acid, 10 mM norepinephrine, and 2 mM L-glutamine (all ingredients from Sigma-Aldrich), at 37 °C in a humid atmosphere of 5% CO2 .

Demethylation Assay:

Article Title: Methylation of a Single Intronic CpG Mediates Expression Silencing of the PMP24 Gene in Prostate Cancer
Article Snippet: .. For the demethylation assay, PC-3 cells were seeded at a density of 1 × 105 cells per T25 flask at day 0, one day before treatment with 5-aza-dC (Sigma, St. Louis, MO). ..

Plasmid Preparation:

Article Title: Fanconi Anemia Gene Editing by the CRISPR/Cas9 System
Article Snippet: PCR products were cloned into the pCR 4 TOPO vector (Invitrogen) for Sanger sequencing confirmation of the entire amplicon using the M13 forward and reverse primers. .. H2AX staining was performed on cells seeded at a concentration of 120,000 total cells in a T25 flask in the presence of 2 m M hydroxyurea (Sigma) for 48 hr using the H2AX phosphorylation assay kit according to the manufacturer's instructions (EMD Millipore, Billerica, MA).

Article Title: An Influenza Virus M2 Protein Specific Chimeric Antigen Receptor Modulates Influenza A/WSN/33 H1N1 Infection In Vivo
Article Snippet: Transduction of Murine Splenocytes The packaging cells (293T/Phoenix Amp) were prepared and transfected as above except that the pColtGalV plasmid was replaced with pHIT123 (MLV-E Env, provided by Prof. R. Debets, University of Rotterdam). .. Splenocytes were seeded at 5x106 /ml in a T25 flask before addition of 2.5 μg/ml Concanavalin A (Calbiochem), 1ng/ml IL-7 (Santa Cruz) and 100 U/ml human rIL-2, and incubated for 24 hours.

Stem Cell Culture:

Article Title: Transgenic Enrichment of Mouse Embryonic Stem Cell-derived Progenitor Motor Neurons
Article Snippet: Paragraph title: 5.1 Embryonic Stem Cell Culture ... Cells were passaged at a 1:5 ratio every 2 days and seeded on a new T25 flask coated with a 0.1% gelatin solution (Sigma).

Transgenic Assay:

Article Title: Multi-Electrode Array with a Planar Surface for Cell Patterning by Microprinting
Article Snippet: HL-1 Cell Culturing, Measurements of Action Potentials, Staining, and Visualization HL-1 cells (cardiac myocytes from transgenic mice) were obtained from Sigma Aldrich. .. HL-1 cells were cultured on the plastic bottom of a T25 flask in Claycomb Medium, supplemented with 10% fetal bovine serum, 100 U/mL penicillin, 100 mg/mL streptomycin, 0.3 mM ascorbic acid, 10 mM norepinephrine, and 2 mM L-glutamine (all ingredients from Sigma-Aldrich), at 37 °C in a humid atmosphere of 5% CO2 .

Phosphorylation Assay:

Article Title: Fanconi Anemia Gene Editing by the CRISPR/Cas9 System
Article Snippet: .. H2AX staining was performed on cells seeded at a concentration of 120,000 total cells in a T25 flask in the presence of 2 m M hydroxyurea (Sigma) for 48 hr using the H2AX phosphorylation assay kit according to the manufacturer's instructions (EMD Millipore, Billerica, MA). .. Flow cytometry was performed using the BD LSRFortessa Cell Analyzer (BD Biosciences).

Concentration Assay:

Article Title: Fanconi Anemia Gene Editing by the CRISPR/Cas9 System
Article Snippet: .. H2AX staining was performed on cells seeded at a concentration of 120,000 total cells in a T25 flask in the presence of 2 m M hydroxyurea (Sigma) for 48 hr using the H2AX phosphorylation assay kit according to the manufacturer's instructions (EMD Millipore, Billerica, MA). .. Flow cytometry was performed using the BD LSRFortessa Cell Analyzer (BD Biosciences).

Article Title: C9orf72 hypermethylation protects against repeat expansion-associated pathology in ALS/FTD
Article Snippet: .. Cells were seeded at a concentration of 300,000 cells/ml in 10 ml in a T25 flask, and 0.5 μM 5-aza-dC (Sigma-Aldrich, St. Louis, MO, USA) was spiked daily into lymphoblast cells for 3 days followed by a 3-day recovery period without 5-aza-dC. ..

Lysis:

Article Title: Palmitoylation of Desmoglein 2 Is a Regulator of Assembly Dynamics and Protein Turnover *
Article Snippet: Cells grown in a T25 flask were scraped into 1 ml of Triton X-100-containing buffer (10 m m Tris-HCl (pH 7.0), 0.5% Triton X-100 (Sigma), 5 m m EDTA, 2 m m EGTA, 30 m m sodium fluoride, 40 m m β-glycerophosphate, 10 m m sodium pyrophosphate, 2 m m sodium orthovanadate, and 2 m m phenylmethylsulfonyl fluoride) and incubated on ice while shaking for 15 min. .. The insoluble pellet was washed once with Triton X-100 lysis buffer prior to resuspension of the pellet in 1 ml of SDS containing buffer (10 m m Tris HCl (pH 8.0), 2% SDS, and 2 m m EDTA).

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    Millipore t25 flasks
    T25 Flasks, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 96 stars, based on 1 article reviews
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    Millipore t25 culture flask
    T25 Culture Flask, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t25 culture flask/product/Millipore
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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