Irradiation:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
Fluorescence:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
Labeling:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
SYBR Green Assay:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
Polymerase Chain Reaction:Article Title: Significant Association Between CAV1 Variant rs3807989 on 7p31 and Atrial Fibrillation in a Chinese Han Population
Article Snippet: .. A total of 25 μL of polymerase chain reaction (PCR) mixture for genotyping contained 1.5 mmol/L of Mg2+ , 0.2 mmol/L of dNTPs, 0.5 μmol/L of each primer, 25 ng of human genomic DNA template, 5 μmol/L of SYTO 9 green fluorescent, and 0.15 U of Taq DNA polymerase (TIANGEN, Beijing, China). .. PCR was performed on an ABI 9700 System (Applied Biosystems, Foster City, CA) with a thermal profile of 95°C for 5 minutes, 40 cycles of 95°C for 10 seconds, 59.4°C or other appropriate annealing temperatures for 10 seconds and 72°C for 15 seconds, and 72°C for 10 minutes.
Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
Reverse Transcription Polymerase Chain Reaction:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
Real-time Polymerase Chain Reaction:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
Plasmid Preparation:Article Title: Induction and Regulation of the Immunoproteasome Subunit β5i (PSMB8) in Laryngeal and Hypopharyngeal Carcinoma Cells
Article Snippet: .. Main reagents The main reagents were Opti-MEM (Thermo Fisher Scientific Inc., USA); polymerase chain reaction (PCR) mix (Tiangen Biotech (Beijing) CO., LTD.); real-time fluorescence quantitative PCR (RT-PCR) mixture and iTaq™ SYBR Green Supermix (Bio-Rad); β5i (LMP7) antibody (Santa Cruz Biotechnology); horseradish peroxidase (HRP) -labeled β-actin mouse monoclonal antibody (Millipore Sigma Canada Co.); HRP-labeled rabbit anti-goat antibody (Amersham Pharmacia Biotech); complementary DNA (cDNA) RT-PCR kit (Promega GoScript™ Reverse Transcription System); γ-ray irradiation source: 60 Co (Beijing Institute of Biotechnology, Beijing 100850, China); plasmid vectors pLKO.1-TRC (Addgene plasmid #10878; http://n2t.net/addgene: 10878; RRID: Addgene_10878), psPAX2, and pMD2. .. G (Invitrogen); DNA restriction endonucleases AgeI (NEB, #R0552S), EcoRI (NEB, #R0101S), and NcoI (NEB, #R3193), along with Buffer 2 (NEB, #B7002S) and T4 DNA Ligase (NEB, #M0202S); FuGENE HD transfection reagent (Roche); molecular weight standard: DNA Marker (DL2000, DL15000) (Tiangen Biotech (Beijing)); low-molecular-weight protein standard (PageRuler Prestained protein ladder, 26616) (Thermo Scientific); GelRed nucleic acid dye (BIOTIUM); DNA fragment extraction kit (A7181, column centrifugation) and PCR product purification kit (Promega); plasmid extraction kit and RNA extraction and purification kit (Tiangen Biotech, Beijing); the exogenous tyrosine kinase inhibitor (TKI) nilotinib (Novartis Pharma Stein AG); human recombinant H-IFN-γ (PeproTech, US), with a final concentration of 150 U/ml; c-Abl activator DPH (MedChemExpress, US), with a final concentration of 10 mM.
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