Purification:Article Title: A replicating recombinant vesicular stomatitis virus model for dairy cattle H5N1 influenza virus glycoprotein evolution
Article Snippet: .. Purified virus was boiled in Laemmeli buffer under reducing conditions and run on a 4-20% acrylamide gel (BioRad). .. Gels were stained with Coomassie protein stain and imaged with a LiCor Odyssey CLx imager (LICORbio).
Virus:Article Title: A replicating recombinant vesicular stomatitis virus model for dairy cattle H5N1 influenza virus glycoprotein evolution
Article Snippet: .. Purified virus was boiled in Laemmeli buffer under reducing conditions and run on a 4-20% acrylamide gel (BioRad). .. Gels were stained with Coomassie protein stain and imaged with a LiCor Odyssey CLx imager (LICORbio).
Acrylamide Gel Assay:Article Title: A replicating recombinant vesicular stomatitis virus model for dairy cattle H5N1 influenza virus glycoprotein evolution
Article Snippet: .. Purified virus was boiled in Laemmeli buffer under reducing conditions and run on a 4-20% acrylamide gel (BioRad). .. Gels were stained with Coomassie protein stain and imaged with a LiCor Odyssey CLx imager (LICORbio).
Lysis:Article Title: Endothelial Notch1 signaling in white adipose tissue promotes cancer cachexia
Article Snippet: .. Cells and tissues in lysis buffer (Cell Signaling) containing 1 mM phenylmethylsulfonyl fluoride were boiled at 95 °C for 5 min in Laemmeli buffer and separated according to molecular weight using SDS–PAGE (Bio-Rad). .. Blotting was performed in a blotting chamber (Peqlab Biotechnology), and proteins were transferred onto nitrocellulose paper.
Article Title: Endothelial Notch1 signaling in white adipose tissue promotes cancer cachexia.
Article Snippet: .. Cells and tissues in lysis buffer (Cell Signaling) containing 1 mM phenylmethylsulfonyl fluoride were boiled at 95 °C for 5 min in Laemmeli buffer and separated according to molecular weight using SDS–PAGE (Bio-Rad). .. Blotting was performed in a blotting chamber (Peqlab Biotechnology), and proteins were transferred onto nitrocellulose paper.
Molecular Weight:Article Title: Endothelial Notch1 signaling in white adipose tissue promotes cancer cachexia
Article Snippet: .. Cells and tissues in lysis buffer (Cell Signaling) containing 1 mM phenylmethylsulfonyl fluoride were boiled at 95 °C for 5 min in Laemmeli buffer and separated according to molecular weight using SDS–PAGE (Bio-Rad). .. Blotting was performed in a blotting chamber (Peqlab Biotechnology), and proteins were transferred onto nitrocellulose paper.
Article Title: Endothelial Notch1 signaling in white adipose tissue promotes cancer cachexia.
Article Snippet: .. Cells and tissues in lysis buffer (Cell Signaling) containing 1 mM phenylmethylsulfonyl fluoride were boiled at 95 °C for 5 min in Laemmeli buffer and separated according to molecular weight using SDS–PAGE (Bio-Rad). .. Blotting was performed in a blotting chamber (Peqlab Biotechnology), and proteins were transferred onto nitrocellulose paper.
SDS Page:Article Title: Plaat1 deficiency reduces cardiac cardiolipin content and impairs exercise tolerance.
Article Snippet: .. Samples were mixed with 6× Laemmeli Buffer (125 mM Tris–HCl, pH 6.8, 20% glycerol (v/v), 4% SDS (w/v), 10% 2-mercaptoethanol (v/v), and 0.05% bromophenol blue) and heated to 95 °C for 5 min, then electrophoresed through 10% SDS-PAGE TGX Stain-FreeTM Fast CastTM gels (Bio-Rad Canada, Mississauga, Ontario, Canada) at 120 V for 1 h. Total loaded protein per lane was detected by fluorescence-activation of the TGX Stain-FreeTM gel under UV illumination, imaged for relative density using a ChemiDOC Touch Imaging System (Bio-Rad Canada), and quantified using Image Lab software (Bio-Rad Canada). .. Proteins were then transferred onto nitrocellulose membranes using a BioRad Trans-Blot Turbo system (Bio-Rad Canada), set at 25 V for 30 min. Membranes were blocked with 5% bovine serum albumin (BSA) in TBST (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4°C in TBST containing, 5% BSA and primary antibodies (1:1000 dilution) directed against heat shock protein 60 (HSP60), succinate dehydrogenase complex subunit A (SDHA), and cytochrome c oxidase subunit IV (COX IV) (Cell Signaling Technology, Whitby, Ontario, Canada).
Article Title: Plaat1 deficiency reduces cardiac cardiolipin content and impairs exercise tolerance
Article Snippet: .. Samples were mixed with 6× Laemmeli Buffer (125 mM Tris–HCl, pH 6.8, 20% glycerol (v/v), 4% SDS (w/v), 10% 2-mercaptoethanol (v/v), and 0.05% bromophenol blue) and heated to 95°C for 5 min, then electrophoresed through 10% SDS-PAGE TGX Stain-FreeTM Fast CastTM gels (Bio-Rad Canada, Mississauga, Ontario, Canada) at 120 V for 1 h. Total loaded protein per lane was detected by fluorescence-activation of the TGX Stain-FreeTM gel under UV illumination, imaged for relative density using a ChemiDOC Touch Imaging System (Bio-Rad Canada), and quantified using Image Lab software (Bio-Rad Canada). .. Proteins were then transferred onto nitrocellulose membranes using a Bio-Rad Trans-Blot Turbo system (Bio-Rad Canada) set at 25 V for 30 min. Membranes were blocked with 5% bovine serum albumin (BSA) in TBST (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4°C in TBST containing 5% BSA and primary antibodies (1:1000 dilution) directed against heat shock protein 60 (HSP60), succinate dehydrogenase complex flavoprotein subunit A (SDHA), and cytochrome c oxidase subunit IV (COX IV) (Cell Signaling Technology).
Staining:Article Title: Plaat1 deficiency reduces cardiac cardiolipin content and impairs exercise tolerance.
Article Snippet: .. Samples were mixed with 6× Laemmeli Buffer (125 mM Tris–HCl, pH 6.8, 20% glycerol (v/v), 4% SDS (w/v), 10% 2-mercaptoethanol (v/v), and 0.05% bromophenol blue) and heated to 95 °C for 5 min, then electrophoresed through 10% SDS-PAGE TGX Stain-FreeTM Fast CastTM gels (Bio-Rad Canada, Mississauga, Ontario, Canada) at 120 V for 1 h. Total loaded protein per lane was detected by fluorescence-activation of the TGX Stain-FreeTM gel under UV illumination, imaged for relative density using a ChemiDOC Touch Imaging System (Bio-Rad Canada), and quantified using Image Lab software (Bio-Rad Canada). .. Proteins were then transferred onto nitrocellulose membranes using a BioRad Trans-Blot Turbo system (Bio-Rad Canada), set at 25 V for 30 min. Membranes were blocked with 5% bovine serum albumin (BSA) in TBST (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4°C in TBST containing, 5% BSA and primary antibodies (1:1000 dilution) directed against heat shock protein 60 (HSP60), succinate dehydrogenase complex subunit A (SDHA), and cytochrome c oxidase subunit IV (COX IV) (Cell Signaling Technology, Whitby, Ontario, Canada).
Article Title: Plaat1 deficiency reduces cardiac cardiolipin content and impairs exercise tolerance
Article Snippet: .. Samples were mixed with 6× Laemmeli Buffer (125 mM Tris–HCl, pH 6.8, 20% glycerol (v/v), 4% SDS (w/v), 10% 2-mercaptoethanol (v/v), and 0.05% bromophenol blue) and heated to 95°C for 5 min, then electrophoresed through 10% SDS-PAGE TGX Stain-FreeTM Fast CastTM gels (Bio-Rad Canada, Mississauga, Ontario, Canada) at 120 V for 1 h. Total loaded protein per lane was detected by fluorescence-activation of the TGX Stain-FreeTM gel under UV illumination, imaged for relative density using a ChemiDOC Touch Imaging System (Bio-Rad Canada), and quantified using Image Lab software (Bio-Rad Canada). .. Proteins were then transferred onto nitrocellulose membranes using a Bio-Rad Trans-Blot Turbo system (Bio-Rad Canada) set at 25 V for 30 min. Membranes were blocked with 5% bovine serum albumin (BSA) in TBST (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4°C in TBST containing 5% BSA and primary antibodies (1:1000 dilution) directed against heat shock protein 60 (HSP60), succinate dehydrogenase complex flavoprotein subunit A (SDHA), and cytochrome c oxidase subunit IV (COX IV) (Cell Signaling Technology).
Fluorescence:Article Title: Plaat1 deficiency reduces cardiac cardiolipin content and impairs exercise tolerance.
Article Snippet: .. Samples were mixed with 6× Laemmeli Buffer (125 mM Tris–HCl, pH 6.8, 20% glycerol (v/v), 4% SDS (w/v), 10% 2-mercaptoethanol (v/v), and 0.05% bromophenol blue) and heated to 95 °C for 5 min, then electrophoresed through 10% SDS-PAGE TGX Stain-FreeTM Fast CastTM gels (Bio-Rad Canada, Mississauga, Ontario, Canada) at 120 V for 1 h. Total loaded protein per lane was detected by fluorescence-activation of the TGX Stain-FreeTM gel under UV illumination, imaged for relative density using a ChemiDOC Touch Imaging System (Bio-Rad Canada), and quantified using Image Lab software (Bio-Rad Canada). .. Proteins were then transferred onto nitrocellulose membranes using a BioRad Trans-Blot Turbo system (Bio-Rad Canada), set at 25 V for 30 min. Membranes were blocked with 5% bovine serum albumin (BSA) in TBST (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4°C in TBST containing, 5% BSA and primary antibodies (1:1000 dilution) directed against heat shock protein 60 (HSP60), succinate dehydrogenase complex subunit A (SDHA), and cytochrome c oxidase subunit IV (COX IV) (Cell Signaling Technology, Whitby, Ontario, Canada).
Article Title: Plaat1 deficiency reduces cardiac cardiolipin content and impairs exercise tolerance
Article Snippet: .. Samples were mixed with 6× Laemmeli Buffer (125 mM Tris–HCl, pH 6.8, 20% glycerol (v/v), 4% SDS (w/v), 10% 2-mercaptoethanol (v/v), and 0.05% bromophenol blue) and heated to 95°C for 5 min, then electrophoresed through 10% SDS-PAGE TGX Stain-FreeTM Fast CastTM gels (Bio-Rad Canada, Mississauga, Ontario, Canada) at 120 V for 1 h. Total loaded protein per lane was detected by fluorescence-activation of the TGX Stain-FreeTM gel under UV illumination, imaged for relative density using a ChemiDOC Touch Imaging System (Bio-Rad Canada), and quantified using Image Lab software (Bio-Rad Canada). .. Proteins were then transferred onto nitrocellulose membranes using a Bio-Rad Trans-Blot Turbo system (Bio-Rad Canada) set at 25 V for 30 min. Membranes were blocked with 5% bovine serum albumin (BSA) in TBST (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4°C in TBST containing 5% BSA and primary antibodies (1:1000 dilution) directed against heat shock protein 60 (HSP60), succinate dehydrogenase complex flavoprotein subunit A (SDHA), and cytochrome c oxidase subunit IV (COX IV) (Cell Signaling Technology).
Imaging:Article Title: Plaat1 deficiency reduces cardiac cardiolipin content and impairs exercise tolerance.
Article Snippet: .. Samples were mixed with 6× Laemmeli Buffer (125 mM Tris–HCl, pH 6.8, 20% glycerol (v/v), 4% SDS (w/v), 10% 2-mercaptoethanol (v/v), and 0.05% bromophenol blue) and heated to 95 °C for 5 min, then electrophoresed through 10% SDS-PAGE TGX Stain-FreeTM Fast CastTM gels (Bio-Rad Canada, Mississauga, Ontario, Canada) at 120 V for 1 h. Total loaded protein per lane was detected by fluorescence-activation of the TGX Stain-FreeTM gel under UV illumination, imaged for relative density using a ChemiDOC Touch Imaging System (Bio-Rad Canada), and quantified using Image Lab software (Bio-Rad Canada). .. Proteins were then transferred onto nitrocellulose membranes using a BioRad Trans-Blot Turbo system (Bio-Rad Canada), set at 25 V for 30 min. Membranes were blocked with 5% bovine serum albumin (BSA) in TBST (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4°C in TBST containing, 5% BSA and primary antibodies (1:1000 dilution) directed against heat shock protein 60 (HSP60), succinate dehydrogenase complex subunit A (SDHA), and cytochrome c oxidase subunit IV (COX IV) (Cell Signaling Technology, Whitby, Ontario, Canada).
Article Title: Plaat1 deficiency reduces cardiac cardiolipin content and impairs exercise tolerance
Article Snippet: .. Samples were mixed with 6× Laemmeli Buffer (125 mM Tris–HCl, pH 6.8, 20% glycerol (v/v), 4% SDS (w/v), 10% 2-mercaptoethanol (v/v), and 0.05% bromophenol blue) and heated to 95°C for 5 min, then electrophoresed through 10% SDS-PAGE TGX Stain-FreeTM Fast CastTM gels (Bio-Rad Canada, Mississauga, Ontario, Canada) at 120 V for 1 h. Total loaded protein per lane was detected by fluorescence-activation of the TGX Stain-FreeTM gel under UV illumination, imaged for relative density using a ChemiDOC Touch Imaging System (Bio-Rad Canada), and quantified using Image Lab software (Bio-Rad Canada). .. Proteins were then transferred onto nitrocellulose membranes using a Bio-Rad Trans-Blot Turbo system (Bio-Rad Canada) set at 25 V for 30 min. Membranes were blocked with 5% bovine serum albumin (BSA) in TBST (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4°C in TBST containing 5% BSA and primary antibodies (1:1000 dilution) directed against heat shock protein 60 (HSP60), succinate dehydrogenase complex flavoprotein subunit A (SDHA), and cytochrome c oxidase subunit IV (COX IV) (Cell Signaling Technology).
Software:Article Title: Plaat1 deficiency reduces cardiac cardiolipin content and impairs exercise tolerance.
Article Snippet: .. Samples were mixed with 6× Laemmeli Buffer (125 mM Tris–HCl, pH 6.8, 20% glycerol (v/v), 4% SDS (w/v), 10% 2-mercaptoethanol (v/v), and 0.05% bromophenol blue) and heated to 95 °C for 5 min, then electrophoresed through 10% SDS-PAGE TGX Stain-FreeTM Fast CastTM gels (Bio-Rad Canada, Mississauga, Ontario, Canada) at 120 V for 1 h. Total loaded protein per lane was detected by fluorescence-activation of the TGX Stain-FreeTM gel under UV illumination, imaged for relative density using a ChemiDOC Touch Imaging System (Bio-Rad Canada), and quantified using Image Lab software (Bio-Rad Canada). .. Proteins were then transferred onto nitrocellulose membranes using a BioRad Trans-Blot Turbo system (Bio-Rad Canada), set at 25 V for 30 min. Membranes were blocked with 5% bovine serum albumin (BSA) in TBST (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4°C in TBST containing, 5% BSA and primary antibodies (1:1000 dilution) directed against heat shock protein 60 (HSP60), succinate dehydrogenase complex subunit A (SDHA), and cytochrome c oxidase subunit IV (COX IV) (Cell Signaling Technology, Whitby, Ontario, Canada).
Article Title: Plaat1 deficiency reduces cardiac cardiolipin content and impairs exercise tolerance
Article Snippet: .. Samples were mixed with 6× Laemmeli Buffer (125 mM Tris–HCl, pH 6.8, 20% glycerol (v/v), 4% SDS (w/v), 10% 2-mercaptoethanol (v/v), and 0.05% bromophenol blue) and heated to 95°C for 5 min, then electrophoresed through 10% SDS-PAGE TGX Stain-FreeTM Fast CastTM gels (Bio-Rad Canada, Mississauga, Ontario, Canada) at 120 V for 1 h. Total loaded protein per lane was detected by fluorescence-activation of the TGX Stain-FreeTM gel under UV illumination, imaged for relative density using a ChemiDOC Touch Imaging System (Bio-Rad Canada), and quantified using Image Lab software (Bio-Rad Canada). .. Proteins were then transferred onto nitrocellulose membranes using a Bio-Rad Trans-Blot Turbo system (Bio-Rad Canada) set at 25 V for 30 min. Membranes were blocked with 5% bovine serum albumin (BSA) in TBST (50 mM Tris–HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) for 1 h at room temperature, then incubated overnight at 4°C in TBST containing 5% BSA and primary antibodies (1:1000 dilution) directed against heat shock protein 60 (HSP60), succinate dehydrogenase complex flavoprotein subunit A (SDHA), and cytochrome c oxidase subunit IV (COX IV) (Cell Signaling Technology).
Bicinchoninic Acid Protein Assay:Article Title: Glial Cell Adhesion Molecule (GlialCAM) Determines Proliferative versus Invasive Cell States in Glioblastoma
Article Snippet: Whole-cell lysates were collected under normal culture conditions by lysing spheres in RIPA buffer (50 mM Tris-HCl pH 8.0, 150 mM NaCl, 1% sodium deoxycholate, 1% Triton X-100, 0.1% SDS, 1% NP-40, and 1 mM EDTA) containing protease and phosphatase inhibitors (catalog #A32955, #A32957, Thermo Fisher Scientific) to obtain soluble protein fractions. .. Total protein was measured by Pierce BCA Protein Assay Kit (catalog #23227, Thermo Fisher Scientific), and then denatured at 95°C for 10min in 4 Laemmeli buffer (catalog #1610747, BIO-RAD) containing 2.5% 2-ME (catalog #M6250, SigmaAldrich). ..
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