Lysis:Article Title: IL-6 is one of the key factors in the formation of gut tissue resident memory T cells from Naïve T cells
Article Snippet: .. Cell lysates were prepared using lysis buffer (20mM TRIS, 100mM NaCl, 1mM EDTA, 1% Triton X-100, 1mM DTT) with protease and phosphatase inhibitors (Thermo Scientific, Waltham, MA). .. 5 μL of each sample were loaded into wells of 10–20% Tris-Glycine SDS gels (Thermo Scientific, Waltham, MA) and run under reducing conditions, followed by transfer onto nitrocellulose membranes (Thermo Scientific, Waltham, MA) using the iBlot2TM system (Invitrogen, IB 1001).
Article Title: Neuroinflammatory stress preferentially impacts synaptic MAPK signaling and mitochondria in excitatory neurons
Article Snippet: .. Equal volumes of bulk brain homogenates and P2 fractions were diluted in 8 M urea lysis buffer (10 mM TRIS, 100 mM NaH2PO4, pH 8.5) with protease and phosphatase inhibitors (ThermoFisher Scientific, 78,446), sonicated 3 times (Pulse 05 05; Amplitude 30%) on ice, and centrifuged at 18,000 × g for 10 min at 4 ̊C. .. Supernatants were collected and total protein concentration was determined by BCA assay (ThermoFisher Scientific, 23,225).
Recombinant:Article Title: Heterocyclic compounds
Article Snippet: .. 1 μL compound dilutions (100% DMSO) were added to 19 μL MAGL (recombinant wild-type) in assay buffer (50 mM TRIS (GIBCO, 15567-027), 1 mM EDTA (Fluka, 03690-100 ml)). .. The plate was shaked for 1 min at 2000 rpm (Variomag Teleshake) and then incubated for 15 min at RT.
Purification:Article Title: Single-Cell and Tissue-Specific CRISPR Editing Analyses Unveil New Insights to Off-Targets and Translocations
Article Snippet: Both procedures were performed according to the Ligation Sequencing gDNA V14 — Whole Genome Amplification protocol (SQK-LSK114, Oxford Nanopore Technologies). .. Minor adjustments were introduced for bead–based purification: the custom buffer was prepared from PEG 8000 (NEB), 0.5 M EDTA (pH 8.0; Thermo Fisher), 5 M NaCl (Sigma–Aldrich), 1 M Tris (pH 8.0; Thermo Fisher), and nuclease–free water (Thermo Fisher). ..
other:
Sonication:Article Title: Neuroinflammatory stress preferentially impacts synaptic MAPK signaling and mitochondria in excitatory neurons
Article Snippet: .. Equal volumes of bulk brain homogenates and P2 fractions were diluted in 8 M urea lysis buffer (10 mM TRIS, 100 mM NaH2PO4, pH 8.5) with protease and phosphatase inhibitors (ThermoFisher Scientific, 78,446), sonicated 3 times (Pulse 05 05; Amplitude 30%) on ice, and centrifuged at 18,000 × g for 10 min at 4 ̊C. .. Supernatants were collected and total protein concentration was determined by BCA assay (ThermoFisher Scientific, 23,225).
Plasmid Preparation:Article Title: NaP-TRAP: A versatile and accessible workflow to dissect principles of translational regulation and mRNA stability
Article Snippet: .. Plasmid templates (see Supporting Information : Table S1 for all sequences) All the plasmid constructs used in this study were cloned in pCS2+ vector which was generously gifted by Antonio Giraldez lab . Nuclease-free water (Sigma-Aldrich, Cat. No. W4502-1L) 1 M TRIS (Thermo Fisher Scientific, Cat. No. 15567027) KAPA HiFi HotStart Polymerase Readymix (Roche, Cat. No. 07958927001) Oligos (see Supporting Information : Table S1) ordered from Sigma or IDT with standard desalting Oligo pools from GenScript and Twist Bioscience Thermal cycler (BIO-RAD, Model No. S1000) DNA Clean & Concentrator (Zymo, Cat. No. D4006) 6X Gel loading dye (New England Biolabs, Cat. No. B7025) EtBr (BIO-RAD Cat. No. 1610433) 1 kb Plus DNA Ladder (New England Biolabs, Cat. No. N3200) TAE buffer (see REAGENTS AND SOLUTIONS ) Agarose gel 1% (wt/vol, see REAGENTS AND SOLUTIONS ) mMessage mMachine SP6 RNA Polymerase kit (Thermo Fisher Scientific, Cat. No. AM1340) DNase I (New England Biolabs, Cat. No. M0303S) RNA Clean & Concentrator-5 (Zymo, Cat. No. R1013) Filter-tips 20 μL, 200 μL and 1,000 μL (RAININ, Cat. Nos. .. Plasmid templates (see Supporting Information : Table S1 for all sequences) All the plasmid constructs used in this study were cloned in pCS2+ vector which was generously gifted by Antonio Giraldez lab . Nuclease-free water (Sigma-Aldrich, Cat. No. W4502-1L) 1 M TRIS (Thermo Fisher Scientific, Cat. No. 15567027) KAPA HiFi HotStart Polymerase Readymix (Roche, Cat. No. 07958927001) Oligos (see Supporting Information : Table S1) ordered from Sigma or IDT with standard desalting Oligo pools from GenScript and Twist Bioscience Thermal cycler (BIO-RAD, Model No. S1000) DNA Clean & Concentrator (Zymo, Cat. No. D4006) 6X Gel loading dye (New England Biolabs, Cat. No. B7025) EtBr (BIO-RAD Cat. No. 1610433) 1 kb Plus DNA Ladder (New England Biolabs, Cat. No. N3200) TAE buffer (see REAGENTS AND SOLUTIONS ) Agarose gel 1% (wt/vol, see REAGENTS AND SOLUTIONS ) mMessage mMachine SP6 RNA Polymerase kit (Thermo Fisher Scientific, Cat. No. AM1340) DNase I (New England Biolabs, Cat. No. M0303S) RNA Clean & Concentrator-5 (Zymo, Cat. No. R1013) Filter-tips 20 μL, 200 μL and 1,000 μL (RAININ, Cat. Nos.
Construct:Article Title: NaP-TRAP: A versatile and accessible workflow to dissect principles of translational regulation and mRNA stability
Article Snippet: .. Plasmid templates (see Supporting Information : Table S1 for all sequences) All the plasmid constructs used in this study were cloned in pCS2+ vector which was generously gifted by Antonio Giraldez lab . Nuclease-free water (Sigma-Aldrich, Cat. No. W4502-1L) 1 M TRIS (Thermo Fisher Scientific, Cat. No. 15567027) KAPA HiFi HotStart Polymerase Readymix (Roche, Cat. No. 07958927001) Oligos (see Supporting Information : Table S1) ordered from Sigma or IDT with standard desalting Oligo pools from GenScript and Twist Bioscience Thermal cycler (BIO-RAD, Model No. S1000) DNA Clean & Concentrator (Zymo, Cat. No. D4006) 6X Gel loading dye (New England Biolabs, Cat. No. B7025) EtBr (BIO-RAD Cat. No. 1610433) 1 kb Plus DNA Ladder (New England Biolabs, Cat. No. N3200) TAE buffer (see REAGENTS AND SOLUTIONS ) Agarose gel 1% (wt/vol, see REAGENTS AND SOLUTIONS ) mMessage mMachine SP6 RNA Polymerase kit (Thermo Fisher Scientific, Cat. No. AM1340) DNase I (New England Biolabs, Cat. No. M0303S) RNA Clean & Concentrator-5 (Zymo, Cat. No. R1013) Filter-tips 20 μL, 200 μL and 1,000 μL (RAININ, Cat. Nos. .. Plasmid templates (see Supporting Information : Table S1 for all sequences) All the plasmid constructs used in this study were cloned in pCS2+ vector which was generously gifted by Antonio Giraldez lab . Nuclease-free water (Sigma-Aldrich, Cat. No. W4502-1L) 1 M TRIS (Thermo Fisher Scientific, Cat. No. 15567027) KAPA HiFi HotStart Polymerase Readymix (Roche, Cat. No. 07958927001) Oligos (see Supporting Information : Table S1) ordered from Sigma or IDT with standard desalting Oligo pools from GenScript and Twist Bioscience Thermal cycler (BIO-RAD, Model No. S1000) DNA Clean & Concentrator (Zymo, Cat. No. D4006) 6X Gel loading dye (New England Biolabs, Cat. No. B7025) EtBr (BIO-RAD Cat. No. 1610433) 1 kb Plus DNA Ladder (New England Biolabs, Cat. No. N3200) TAE buffer (see REAGENTS AND SOLUTIONS ) Agarose gel 1% (wt/vol, see REAGENTS AND SOLUTIONS ) mMessage mMachine SP6 RNA Polymerase kit (Thermo Fisher Scientific, Cat. No. AM1340) DNase I (New England Biolabs, Cat. No. M0303S) RNA Clean & Concentrator-5 (Zymo, Cat. No. R1013) Filter-tips 20 μL, 200 μL and 1,000 μL (RAININ, Cat. Nos.
Clone Assay:Article Title: NaP-TRAP: A versatile and accessible workflow to dissect principles of translational regulation and mRNA stability
Article Snippet: .. Plasmid templates (see Supporting Information : Table S1 for all sequences) All the plasmid constructs used in this study were cloned in pCS2+ vector which was generously gifted by Antonio Giraldez lab . Nuclease-free water (Sigma-Aldrich, Cat. No. W4502-1L) 1 M TRIS (Thermo Fisher Scientific, Cat. No. 15567027) KAPA HiFi HotStart Polymerase Readymix (Roche, Cat. No. 07958927001) Oligos (see Supporting Information : Table S1) ordered from Sigma or IDT with standard desalting Oligo pools from GenScript and Twist Bioscience Thermal cycler (BIO-RAD, Model No. S1000) DNA Clean & Concentrator (Zymo, Cat. No. D4006) 6X Gel loading dye (New England Biolabs, Cat. No. B7025) EtBr (BIO-RAD Cat. No. 1610433) 1 kb Plus DNA Ladder (New England Biolabs, Cat. No. N3200) TAE buffer (see REAGENTS AND SOLUTIONS ) Agarose gel 1% (wt/vol, see REAGENTS AND SOLUTIONS ) mMessage mMachine SP6 RNA Polymerase kit (Thermo Fisher Scientific, Cat. No. AM1340) DNase I (New England Biolabs, Cat. No. M0303S) RNA Clean & Concentrator-5 (Zymo, Cat. No. R1013) Filter-tips 20 μL, 200 μL and 1,000 μL (RAININ, Cat. Nos. .. Plasmid templates (see Supporting Information : Table S1 for all sequences) All the plasmid constructs used in this study were cloned in pCS2+ vector which was generously gifted by Antonio Giraldez lab . Nuclease-free water (Sigma-Aldrich, Cat. No. W4502-1L) 1 M TRIS (Thermo Fisher Scientific, Cat. No. 15567027) KAPA HiFi HotStart Polymerase Readymix (Roche, Cat. No. 07958927001) Oligos (see Supporting Information : Table S1) ordered from Sigma or IDT with standard desalting Oligo pools from GenScript and Twist Bioscience Thermal cycler (BIO-RAD, Model No. S1000) DNA Clean & Concentrator (Zymo, Cat. No. D4006) 6X Gel loading dye (New England Biolabs, Cat. No. B7025) EtBr (BIO-RAD Cat. No. 1610433) 1 kb Plus DNA Ladder (New England Biolabs, Cat. No. N3200) TAE buffer (see REAGENTS AND SOLUTIONS ) Agarose gel 1% (wt/vol, see REAGENTS AND SOLUTIONS ) mMessage mMachine SP6 RNA Polymerase kit (Thermo Fisher Scientific, Cat. No. AM1340) DNase I (New England Biolabs, Cat. No. M0303S) RNA Clean & Concentrator-5 (Zymo, Cat. No. R1013) Filter-tips 20 μL, 200 μL and 1,000 μL (RAININ, Cat. Nos.
Agarose Gel Electrophoresis:Article Title: NaP-TRAP: A versatile and accessible workflow to dissect principles of translational regulation and mRNA stability
Article Snippet: .. Plasmid templates (see Supporting Information : Table S1 for all sequences) All the plasmid constructs used in this study were cloned in pCS2+ vector which was generously gifted by Antonio Giraldez lab . Nuclease-free water (Sigma-Aldrich, Cat. No. W4502-1L) 1 M TRIS (Thermo Fisher Scientific, Cat. No. 15567027) KAPA HiFi HotStart Polymerase Readymix (Roche, Cat. No. 07958927001) Oligos (see Supporting Information : Table S1) ordered from Sigma or IDT with standard desalting Oligo pools from GenScript and Twist Bioscience Thermal cycler (BIO-RAD, Model No. S1000) DNA Clean & Concentrator (Zymo, Cat. No. D4006) 6X Gel loading dye (New England Biolabs, Cat. No. B7025) EtBr (BIO-RAD Cat. No. 1610433) 1 kb Plus DNA Ladder (New England Biolabs, Cat. No. N3200) TAE buffer (see REAGENTS AND SOLUTIONS ) Agarose gel 1% (wt/vol, see REAGENTS AND SOLUTIONS ) mMessage mMachine SP6 RNA Polymerase kit (Thermo Fisher Scientific, Cat. No. AM1340) DNase I (New England Biolabs, Cat. No. M0303S) RNA Clean & Concentrator-5 (Zymo, Cat. No. R1013) Filter-tips 20 μL, 200 μL and 1,000 μL (RAININ, Cat. Nos. .. Plasmid templates (see Supporting Information : Table S1 for all sequences) All the plasmid constructs used in this study were cloned in pCS2+ vector which was generously gifted by Antonio Giraldez lab . Nuclease-free water (Sigma-Aldrich, Cat. No. W4502-1L) 1 M TRIS (Thermo Fisher Scientific, Cat. No. 15567027) KAPA HiFi HotStart Polymerase Readymix (Roche, Cat. No. 07958927001) Oligos (see Supporting Information : Table S1) ordered from Sigma or IDT with standard desalting Oligo pools from GenScript and Twist Bioscience Thermal cycler (BIO-RAD, Model No. S1000) DNA Clean & Concentrator (Zymo, Cat. No. D4006) 6X Gel loading dye (New England Biolabs, Cat. No. B7025) EtBr (BIO-RAD Cat. No. 1610433) 1 kb Plus DNA Ladder (New England Biolabs, Cat. No. N3200) TAE buffer (see REAGENTS AND SOLUTIONS ) Agarose gel 1% (wt/vol, see REAGENTS AND SOLUTIONS ) mMessage mMachine SP6 RNA Polymerase kit (Thermo Fisher Scientific, Cat. No. AM1340) DNase I (New England Biolabs, Cat. No. M0303S) RNA Clean & Concentrator-5 (Zymo, Cat. No. R1013) Filter-tips 20 μL, 200 μL and 1,000 μL (RAININ, Cat. Nos.
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