Extraction:Article Title: IGDQ motogenic peptide gradient induces directional cell migration through integrin (αv)β3 activation in MDA-MB-231 metastatic breast cancer cells.
Article Snippet: The medium was then replaced by fresh complete medium ontaining puromycin (2 μg/mL – Sigma-Aldrich) to select transduced cells. he selection was maintained during the following experiments. .. NA extraction MDA-MB-231 cells grown in T25 flask (0.4 ∗ 10 6 cells for 48h) were ashed with PBS (20 mL phosphate buffer - KH 2 PO 4 0.5 mM at pH 7.4, 9g aCl qsp 1L bidistilled water) on ice, scraped off the plastic surface in PBS, entrifuged in 1.5 mL Eppendorf tube at 1000 rpm – 4 °C and cell pellet arvested with 600 μL of RLT Lysis Buffer from RNeasy Mini Kit (Qiagen) n a 2 mL tube. .. Total RNA was extracted using the RNeasy Mini Kit and he automate QIAcube (Qiagen, Germany) under conditions “Large sample ith DNase” and an elution volume of 30 μL.
Multiple Displacement Amplification:Article Title: IGDQ motogenic peptide gradient induces directional cell migration through integrin (αv)β3 activation in MDA-MB-231 metastatic breast cancer cells.
Article Snippet: The medium was then replaced by fresh complete medium ontaining puromycin (2 μg/mL – Sigma-Aldrich) to select transduced cells. he selection was maintained during the following experiments. .. NA extraction MDA-MB-231 cells grown in T25 flask (0.4 ∗ 10 6 cells for 48h) were ashed with PBS (20 mL phosphate buffer - KH 2 PO 4 0.5 mM at pH 7.4, 9g aCl qsp 1L bidistilled water) on ice, scraped off the plastic surface in PBS, entrifuged in 1.5 mL Eppendorf tube at 1000 rpm – 4 °C and cell pellet arvested with 600 μL of RLT Lysis Buffer from RNeasy Mini Kit (Qiagen) n a 2 mL tube. .. Total RNA was extracted using the RNeasy Mini Kit and he automate QIAcube (Qiagen, Germany) under conditions “Large sample ith DNase” and an elution volume of 30 μL.
Article Title: Indacaterol inhibits collective cell migration and IGDQ-mediated single cell migration in metastatic breast cancer MDA-MB-231 cells
Article Snippet: Indacaterol maleate 15μM (stock solution at 20 mM in DMSO – Sigma Aldrich / Merck, Germany ) was used to treat cells 24h after cell seeding, for 24h, 48h or 5 days. .. MDA-MB-231 cells grown in T25 flask (0.4*10 6 cells for 48h) were washed with PBS (20 mL phosphate buffer—KH 2 PO 4 0.5 mM at pH 7.4, 9 g NaCl qsp 1L bidistilled water) on ice, scraped off the plastic surface in PBS, centrifuged in 1.5 mL Eppendorf tube at 1000 rpm – 4°C and cell pellet harvested with 600 μL of RLT Lysis Buffer from RNeasy Mini Kit ( Qiagen, Germany ) in a 2 mL tube. .. Total RNA was extracted using the RNeasy Mini Kit and the automate QIAcube ( Qiagen, Germany ) under conditions “Large sample with DNase” and an elution volume of 30 μL.
Article Title: Indacaterol inhibits collective cell migration and IGDQ-mediated single cell migration in metastatic breast cancer MDA-MB-231 cells.
Article Snippet: Indacaterol maleate 15μM (stock solution at 20 mM in DMSO – Sigma Aldrich / Merck, Germany) was used to treat cells 24h after cell seeding, for 24h, 48h or 5 days. .. MDA-MB-231 cells grown in T25 flask (0.4*106 cells for 48h) were washed with PBS (20 mL phosphate buffer— KH2PO4 0.5 mM at pH 7.4, 9 g NaCl qsp 1L bidistilled water) on ice, scraped off the plastic surface in PBS, centrifuged in 1.5 mL Eppendorf tube at 1000 rpm – 4°C and cell pellet harvested with 600 μL of RLT Lysis Buffer from RNeasy Mini Kit (Qiagen, Germany) in a 2 mL tube. .. Total RNA was extracted using the RNeasy Mini Kit and the automate QIAcube (Qiagen, Germany) under conditions “Large sample with DNase” and an elution volume of 30 μL.
Article Title: Indacaterol inhibits collective cell migration and IGDQ-mediated single cell migration in metastatic breast cancer MDA-MB-231 cells
Article Snippet: .. MDA-MB-231 cells grown in T25 flask (0.4*10 6 cells for 48h) were washed with PBS on ice, scraped off the plastic surface in PBS, centrifuged in 1.5 mL Eppendorf tube at 1000 rpm – 4°C and cell pellet harvested with 30 μL of transmembrane protein lysis buffer (stock: 40 mM Trizma base, 150 mM KCl, 2 mM EDTA; lysis buffer: 1 mL stock lysis buffer, 30 μL Phosphatase Inhibitor Cocktail (PIC, Roche), 30 μL Phosphatase Inhibitor Buffer (PIB: 25 mM Na 3 VO 4 , 250 mM Para-NitroPhenyl Phosphate, 250 mM β-glycerophosphate, 125 nM NaF), 2 μL of β-mercaptoethanol 0.2%). ..
Article Title: Indacaterol inhibits collective cell migration and IGDQ-mediated single cell migration in metastatic breast cancer MDA-MB-231 cells.
Article Snippet: .. MDA-MB-231 cells grown in T25 flask (0.4*106 cells for 48h) were washed with PBS on ice, scraped off the plastic surface in PBS, centrifuged in 1.5 mL Eppendorf tube at 1000 rpm – 4°C and cell pellet harvested with 30 μL of transmembrane protein lysis buffer (stock: 40 mM Trizma base, 150 mM KCl, 2 mM EDTA; lysis buffer: 1 mL stock lysis buffer, 30 μL Phosphatase Inhibitor Cocktail (PIC, Roche), 30 μL Phosphatase Inhibitor Buffer (PIB: 25 mM Na3VO4, 250 mM Para-NitroPhenyl Phosphate, 250 mM β-glycerophosphate, 125 nM NaF), 2 μL of β-mercaptoethanol 0.2%). ..
Lysis:Article Title: IGDQ motogenic peptide gradient induces directional cell migration through integrin (αv)β3 activation in MDA-MB-231 metastatic breast cancer cells.
Article Snippet: The medium was then replaced by fresh complete medium ontaining puromycin (2 μg/mL – Sigma-Aldrich) to select transduced cells. he selection was maintained during the following experiments. .. NA extraction MDA-MB-231 cells grown in T25 flask (0.4 ∗ 10 6 cells for 48h) were ashed with PBS (20 mL phosphate buffer - KH 2 PO 4 0.5 mM at pH 7.4, 9g aCl qsp 1L bidistilled water) on ice, scraped off the plastic surface in PBS, entrifuged in 1.5 mL Eppendorf tube at 1000 rpm – 4 °C and cell pellet arvested with 600 μL of RLT Lysis Buffer from RNeasy Mini Kit (Qiagen) n a 2 mL tube. .. Total RNA was extracted using the RNeasy Mini Kit and he automate QIAcube (Qiagen, Germany) under conditions “Large sample ith DNase” and an elution volume of 30 μL.
Article Title: Indacaterol inhibits collective cell migration and IGDQ-mediated single cell migration in metastatic breast cancer MDA-MB-231 cells
Article Snippet: Indacaterol maleate 15μM (stock solution at 20 mM in DMSO – Sigma Aldrich / Merck, Germany ) was used to treat cells 24h after cell seeding, for 24h, 48h or 5 days. .. MDA-MB-231 cells grown in T25 flask (0.4*10 6 cells for 48h) were washed with PBS (20 mL phosphate buffer—KH 2 PO 4 0.5 mM at pH 7.4, 9 g NaCl qsp 1L bidistilled water) on ice, scraped off the plastic surface in PBS, centrifuged in 1.5 mL Eppendorf tube at 1000 rpm – 4°C and cell pellet harvested with 600 μL of RLT Lysis Buffer from RNeasy Mini Kit ( Qiagen, Germany ) in a 2 mL tube. .. Total RNA was extracted using the RNeasy Mini Kit and the automate QIAcube ( Qiagen, Germany ) under conditions “Large sample with DNase” and an elution volume of 30 μL.
Article Title: Indacaterol inhibits collective cell migration and IGDQ-mediated single cell migration in metastatic breast cancer MDA-MB-231 cells.
Article Snippet: Indacaterol maleate 15μM (stock solution at 20 mM in DMSO – Sigma Aldrich / Merck, Germany) was used to treat cells 24h after cell seeding, for 24h, 48h or 5 days. .. MDA-MB-231 cells grown in T25 flask (0.4*106 cells for 48h) were washed with PBS (20 mL phosphate buffer— KH2PO4 0.5 mM at pH 7.4, 9 g NaCl qsp 1L bidistilled water) on ice, scraped off the plastic surface in PBS, centrifuged in 1.5 mL Eppendorf tube at 1000 rpm – 4°C and cell pellet harvested with 600 μL of RLT Lysis Buffer from RNeasy Mini Kit (Qiagen, Germany) in a 2 mL tube. .. Total RNA was extracted using the RNeasy Mini Kit and the automate QIAcube (Qiagen, Germany) under conditions “Large sample with DNase” and an elution volume of 30 μL.
Article Title: Indacaterol inhibits collective cell migration and IGDQ-mediated single cell migration in metastatic breast cancer MDA-MB-231 cells
Article Snippet: .. MDA-MB-231 cells grown in T25 flask (0.4*10 6 cells for 48h) were washed with PBS on ice, scraped off the plastic surface in PBS, centrifuged in 1.5 mL Eppendorf tube at 1000 rpm – 4°C and cell pellet harvested with 30 μL of transmembrane protein lysis buffer (stock: 40 mM Trizma base, 150 mM KCl, 2 mM EDTA; lysis buffer: 1 mL stock lysis buffer, 30 μL Phosphatase Inhibitor Cocktail (PIC, Roche), 30 μL Phosphatase Inhibitor Buffer (PIB: 25 mM Na 3 VO 4 , 250 mM Para-NitroPhenyl Phosphate, 250 mM β-glycerophosphate, 125 nM NaF), 2 μL of β-mercaptoethanol 0.2%). ..
Article Title: Indacaterol inhibits collective cell migration and IGDQ-mediated single cell migration in metastatic breast cancer MDA-MB-231 cells.
Article Snippet: .. MDA-MB-231 cells grown in T25 flask (0.4*106 cells for 48h) were washed with PBS on ice, scraped off the plastic surface in PBS, centrifuged in 1.5 mL Eppendorf tube at 1000 rpm – 4°C and cell pellet harvested with 30 μL of transmembrane protein lysis buffer (stock: 40 mM Trizma base, 150 mM KCl, 2 mM EDTA; lysis buffer: 1 mL stock lysis buffer, 30 μL Phosphatase Inhibitor Cocktail (PIC, Roche), 30 μL Phosphatase Inhibitor Buffer (PIB: 25 mM Na3VO4, 250 mM Para-NitroPhenyl Phosphate, 250 mM β-glycerophosphate, 125 nM NaF), 2 μL of β-mercaptoethanol 0.2%). ..
Bioprocessing:Article Title: Efficient Gene-Editing in Human Pluripotent Stem Cells Through Simplified Assembly of Adeno-Associated Viral (AAV) Donor Templates
Article Snippet: .. Co-transfect the AAV-293 cells with the AAV targeting vector along with pHelper and RC-DJ plasmids using the Cal-Phos method: Per T25 flask, prepare in a 1.5 mL Eppendorf tube 37 μL of 2 M CaCl2 solution, 3 μg of each plasmid, and water to a total volume of 300 μL. ..
Plasmid Preparation:Article Title: Efficient Gene-Editing in Human Pluripotent Stem Cells Through Simplified Assembly of Adeno-Associated Viral (AAV) Donor Templates
Article Snippet: .. Co-transfect the AAV-293 cells with the AAV targeting vector along with pHelper and RC-DJ plasmids using the Cal-Phos method: Per T25 flask, prepare in a 1.5 mL Eppendorf tube 37 μL of 2 M CaCl2 solution, 3 μg of each plasmid, and water to a total volume of 300 μL. ..
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