nupage lds sample buffer  (Thermo Fisher)


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    Structured Review

    Thermo Fisher nupage lds sample buffer
    Nupage Lds Sample Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 729 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nupage lds sample buffer/product/Thermo Fisher
    Average 99 stars, based on 729 article reviews
    Price from $9.99 to $1999.99
    nupage lds sample buffer - by Bioz Stars, 2020-03
    99/100 stars

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    Centrifugation:

    Article Title: Lentiviral Vector Production Titer Is Not Limited in HEK293T by Induced Intracellular Innate Immunity
    Article Snippet: .. Whole cell lysis was carried out on ice for 15 min and lysates cleared by centrifugation in a tabletop centrifuge at full speed for 5 min. Total protein was quantified using BCA Protein Assay Reagents (Pierce Thermo Fisher Scientific) and 25 μg total protein extract was heated at 95°C for 5 min with NuPAGE LDS Sample Buffer (Thermo Fisher Scientific) and then loaded on a Novex NuPAGE 4%–12% Bis-Tris gel (Thermo Fisher Scientific). .. Wet transfer was performed using the X-Cell SureLock Blot module (Thermo Fisher Scientific) onto Immun-blot PVDF Membrane (BioRad, Hercules, CA, USA).

    Article Title: The long noncoding RNA NEAT1 and nuclear paraspeckles are up-regulated by the transcription factor HSF1 in the heat shock response
    Article Snippet: Whole-cell extracts (WCE) were made by lysing cells directly in 2× NuPAGE LDS Sample Buffer (ThermoFisher Scientific). .. In brief, cells were resuspended in Cytoplasmic Extraction Reagent I and II, and nuclei were pelleted by centrifugation at 16,000 × g .

    Expressing:

    Article Title: The AKT isoforms 1 and 2 drive B cell fate decisions during the germinal center response
    Article Snippet: For analysis of protein expression in whole cells, whole cell lysates were prepared by direct lysing with radio immunoprecipitation assay buffer plus complete protease inhibitor mixture (Roche). .. The samples were boiled in NuPAGE LDS Sample Buffer (Invitrogen) supplemented with β-mercaptoethanol and then were separated by 4–12% polyacrylamide Bis-Tris gel (Bio-Rad) and transferred onto polyvinylidene difluoride membrane (EMD Millipore).

    Article Title: Bleomycin hydrolase regulates the release of chemokines important for inflammation and wound healing by keratinocytes
    Article Snippet: .. Western blot For Western blot analysis of protein expression, 30 ug of total protein lysates was mixed with NuPAGE LDS Sample buffer (Invitrogen) and NuPAGE Sample Reducing Agent (Invitrogen) and heated for 10 minutes, 70 °C. .. The samples were loaded onto NuPAGE 4–12% Bis-Tris Protein Gels (Invitrogen) and run with NuPAGE MOPS SDS Running buffer (Invitrogen) according to the NuPAGE Novex electrophoresis program.

    Blocking Assay:

    Article Title: Efficacy of Multi-exon Skipping Treatment in Duchenne Muscular Dystrophy Dog Model Neonates
    Article Snippet: Protein was prepared by adding NuPAGE LDS Sample Buffer (Thermo Fisher) and NuPAGE Sample Reducing Agent (Thermo Fisher) at 1× final concentrations into the samples and then incubating at 70°C for 10 min. Western blotting was performed as described previously. .. The primary antibodies used, diluted using 2% Amersham ECL Prime blocking reagent (GE Healthcare) in PBS with 0.05% Tween 20 (PBST), were as follows: NCL-DYS1 (1:200, Leica Biosystems) for the dystrophin rod domain and desmin (1:4,000, Abcam) as a loading control.

    Article Title: TGFβ1‐induced cell motility but not cell proliferation is mediated through Cten in colorectal cancer
    Article Snippet: Fifty micrograms of protein was added to NUPAGE LDS Sample Buffer (Thermo Fisher Scientific) containing 5% β‐mercaptoethanol. .. The protein samples were heated to 90°C on a heat block for 5 minutes and cooled on ice for another 5 minutes.

    Article Title: TGFβ1‐induced cell motility but not cell proliferation is mediated through Cten in colorectal cancer
    Article Snippet: Fifty micrograms of protein was added to NUPAGE LDS Sample Buffer (Thermo Fisher Scientific) containing 5% β‐mercaptoethanol. .. The protein samples were heated to 90°C on a heat block for 5 minutes and cooled on ice for another 5 minutes.

    Article Title: Bleomycin hydrolase regulates the release of chemokines important for inflammation and wound healing by keratinocytes
    Article Snippet: Western blot For Western blot analysis of protein expression, 30 ug of total protein lysates was mixed with NuPAGE LDS Sample buffer (Invitrogen) and NuPAGE Sample Reducing Agent (Invitrogen) and heated for 10 minutes, 70 °C. .. The proteins were transferred to Nitrocellulose Blotting Membranes (Invitrogen) using NuPAGE Transfer buffer (Invitrogen) containing 20% methanol, followed by blocking with 5% milk in PBS + Tween for 1 hour on shaker.

    Article Title: Efficacy of Multi-exon Skipping Treatment in Duchenne Muscular Dystrophy Dog Model Neonates
    Article Snippet: Protein was prepared by adding NuPAGE LDS Sample Buffer (Thermo Fisher) and NuPAGE Sample Reducing Agent (Thermo Fisher) at 1× final concentrations into the samples and then incubating at 70°C for 10 min. Western blotting was performed as described previously. .. The primary antibodies used, diluted using 2% Amersham ECL Prime blocking reagent (GE Healthcare) in PBS with 0.05% Tween 20 (PBST), were as follows: NCL-DYS1 (1:200, Leica Biosystems) for the dystrophin rod domain and desmin (1:4,000, Abcam) as a loading control.

    Suction Filtration:

    Article Title: CAT tails drive degradation of stalled polypeptides on and off the ribosome
    Article Snippet: TEV treatment 1-liter log-phase yeast cultures at OD600 = 0.6–0.8 were harvested by vacuum filtration and flash frozen in liquid nitrogen. .. 4x NuPage LDS Sample Buffer (Thermo Fisher Scientific) with 5% β-mercaptoethanol was added at 1:1 vol:vol before boiling at 95°C for 5 min to denature the samples.

    Electrophoresis:

    Article Title: Bleomycin hydrolase regulates the release of chemokines important for inflammation and wound healing by keratinocytes
    Article Snippet: Western blot For Western blot analysis of protein expression, 30 ug of total protein lysates was mixed with NuPAGE LDS Sample buffer (Invitrogen) and NuPAGE Sample Reducing Agent (Invitrogen) and heated for 10 minutes, 70 °C. .. The samples were loaded onto NuPAGE 4–12% Bis-Tris Protein Gels (Invitrogen) and run with NuPAGE MOPS SDS Running buffer (Invitrogen) according to the NuPAGE Novex electrophoresis program.

    Incubation:

    Article Title: p53 controls genomic stability and temporal differentiation of human neural stem cells and affects neural organization in human brain organoids
    Article Snippet: .. Western blot Whole-cell lysates were prepared by lysing cell pellets in 1×NuPAGE LDS Sample buffer including protease and phosphatase inhibitor (ThermoFisher Scientific, 78440), incubated 5 min at 90 °C and 3 rounds of sonication for 5 min. Total protein was loaded on a 4–12% gradient Bis/Tris gel (ThermoFisher Scientific, NP0322BOX). .. Proteins were transferred to a nitrocellulose membrane, using the Trans-Blot Turbo Transfer System (Bio-Rad, 1704150).

    Article Title: Lentiviral Vector Production Titer Is Not Limited in HEK293T by Induced Intracellular Innate Immunity
    Article Snippet: Whole cell lysis was carried out on ice for 15 min and lysates cleared by centrifugation in a tabletop centrifuge at full speed for 5 min. Total protein was quantified using BCA Protein Assay Reagents (Pierce Thermo Fisher Scientific) and 25 μg total protein extract was heated at 95°C for 5 min with NuPAGE LDS Sample Buffer (Thermo Fisher Scientific) and then loaded on a Novex NuPAGE 4%–12% Bis-Tris gel (Thermo Fisher Scientific). .. Membranes were blocked with 5% skimmed milk in PBS-Tween for 1 h at room temperature and incubated overnight at 4°C with the primary antibody.

    Article Title: Efficacy of Multi-exon Skipping Treatment in Duchenne Muscular Dystrophy Dog Model Neonates
    Article Snippet: This was mixed, incubated at 37°C for 5 min, and spun at max speed for 30 min at 16°C to collect the protein-containing supernatant. .. Protein was prepared by adding NuPAGE LDS Sample Buffer (Thermo Fisher) and NuPAGE Sample Reducing Agent (Thermo Fisher) at 1× final concentrations into the samples and then incubating at 70°C for 10 min. Western blotting was performed as described previously.

    Article Title: Bleomycin hydrolase regulates the release of chemokines important for inflammation and wound healing by keratinocytes
    Article Snippet: Western blot For Western blot analysis of protein expression, 30 ug of total protein lysates was mixed with NuPAGE LDS Sample buffer (Invitrogen) and NuPAGE Sample Reducing Agent (Invitrogen) and heated for 10 minutes, 70 °C. .. For detection of BLMH, the membranes were incubated cold overnight with Human BLMH Antibody (R & D Systems) 1:1000 dilution in blocking buffer.

    Article Title: Autophagy and mitochondrial biogenesis impairment contribute to age-dependent liver injury in experimental sepsis: dysregulation of AMP-activated protein kinase pathway
    Article Snippet: Cytosol and nuclear extracts were boiled in equal volumes of NuPAGE LDS Sample Buffer (Thermo Fisher Scientific) and 25–40 μg of protein loaded per lane on a 16% Tris-glycine gradient gel. .. Membranes were washed in Tris-buffered saline with 0.1% Tween 20 and incubated with secondary peroxidase-conjugated antibody and reprobed with primary antibody against β-actin, to ensure equal loading samples of both cytosol and nuclear proteins ( ).

    Article Title: Protein kinase C epsilon contributes to basal and sensitizing responses of TRPV1 to capsaicin in rat dorsal root ganglion neurons
    Article Snippet: Vector-infected U2OS cells (MOI 3) were harvested at 16 h post-infection (hpi) with 1× NuPAGE LDS Sample Buffer in PBS (Invitrogen, Carlsbad, CA, USA) and heated at 70°C for 10 min. .. Following transfer, the membrane was incubated with anti-GFP goat polyclonal antibody (Abcam, Cambridge, MA, USA) at 1 : 500 dilution in PBS with 3% milk.

    Article Title: Efficacy of Multi-exon Skipping Treatment in Duchenne Muscular Dystrophy Dog Model Neonates
    Article Snippet: This was mixed, incubated at 37°C for 5 min, and spun at max speed for 30 min at 16°C to collect the protein-containing supernatant. .. Protein was prepared by adding NuPAGE LDS Sample Buffer (Thermo Fisher) and NuPAGE Sample Reducing Agent (Thermo Fisher) at 1× final concentrations into the samples and then incubating at 70°C for 10 min. Western blotting was performed as described previously.

    Mass Spectrometry:

    Article Title: The long noncoding RNA neuroLNC regulates presynaptic activity by interacting with the neurodegeneration-associated protein TDP-43
    Article Snippet: .. MS identification of protein interactors following ChIRP/RNA interactome analysis For proteomic analysis, we initially reversed the cross-linking by boiling the beads in a final concentration of 1× NuPAGE LDS Sample Buffer (Thermo Fisher Scientific) at 95°C for 30 min on a thermomixer (at 750 rpm). .. After boiling, the samples were loaded on precast NuPAGE 4 to 12% Bis-Tris Protein Gels (Thermo Fisher Scientific) and separated for 20 min at constant voltage (200 V).

    BIA-KA:

    Article Title: The AKT isoforms 1 and 2 drive B cell fate decisions during the germinal center response
    Article Snippet: Protein concentration of subcellular fractionation and whole cell lysates was quantitated using the BCA protein assay kit (Pierce). .. The samples were boiled in NuPAGE LDS Sample Buffer (Invitrogen) supplemented with β-mercaptoethanol and then were separated by 4–12% polyacrylamide Bis-Tris gel (Bio-Rad) and transferred onto polyvinylidene difluoride membrane (EMD Millipore).

    Article Title: Lentiviral Vector Production Titer Is Not Limited in HEK293T by Induced Intracellular Innate Immunity
    Article Snippet: .. Whole cell lysis was carried out on ice for 15 min and lysates cleared by centrifugation in a tabletop centrifuge at full speed for 5 min. Total protein was quantified using BCA Protein Assay Reagents (Pierce Thermo Fisher Scientific) and 25 μg total protein extract was heated at 95°C for 5 min with NuPAGE LDS Sample Buffer (Thermo Fisher Scientific) and then loaded on a Novex NuPAGE 4%–12% Bis-Tris gel (Thermo Fisher Scientific). .. Wet transfer was performed using the X-Cell SureLock Blot module (Thermo Fisher Scientific) onto Immun-blot PVDF Membrane (BioRad, Hercules, CA, USA).

    Western Blot:

    Article Title: The AKT isoforms 1 and 2 drive B cell fate decisions during the germinal center response
    Article Snippet: Paragraph title: Subcellular fractionation and Western blot analysis ... The samples were boiled in NuPAGE LDS Sample Buffer (Invitrogen) supplemented with β-mercaptoethanol and then were separated by 4–12% polyacrylamide Bis-Tris gel (Bio-Rad) and transferred onto polyvinylidene difluoride membrane (EMD Millipore).

    Article Title: p53 controls genomic stability and temporal differentiation of human neural stem cells and affects neural organization in human brain organoids
    Article Snippet: .. Western blot Whole-cell lysates were prepared by lysing cell pellets in 1×NuPAGE LDS Sample buffer including protease and phosphatase inhibitor (ThermoFisher Scientific, 78440), incubated 5 min at 90 °C and 3 rounds of sonication for 5 min. Total protein was loaded on a 4–12% gradient Bis/Tris gel (ThermoFisher Scientific, NP0322BOX). .. Proteins were transferred to a nitrocellulose membrane, using the Trans-Blot Turbo Transfer System (Bio-Rad, 1704150).

    Article Title: Lentiviral Vector Production Titer Is Not Limited in HEK293T by Induced Intracellular Innate Immunity
    Article Snippet: Paragraph title: Immunoblots ... Whole cell lysis was carried out on ice for 15 min and lysates cleared by centrifugation in a tabletop centrifuge at full speed for 5 min. Total protein was quantified using BCA Protein Assay Reagents (Pierce Thermo Fisher Scientific) and 25 μg total protein extract was heated at 95°C for 5 min with NuPAGE LDS Sample Buffer (Thermo Fisher Scientific) and then loaded on a Novex NuPAGE 4%–12% Bis-Tris gel (Thermo Fisher Scientific).

    Article Title: Efficacy of Multi-exon Skipping Treatment in Duchenne Muscular Dystrophy Dog Model Neonates
    Article Snippet: .. Protein was prepared by adding NuPAGE LDS Sample Buffer (Thermo Fisher) and NuPAGE Sample Reducing Agent (Thermo Fisher) at 1× final concentrations into the samples and then incubating at 70°C for 10 min. Western blotting was performed as described previously. ..

    Article Title: TGFβ1‐induced cell motility but not cell proliferation is mediated through Cten in colorectal cancer
    Article Snippet: Paragraph title: 2.4. Western blot ... Fifty micrograms of protein was added to NUPAGE LDS Sample Buffer (Thermo Fisher Scientific) containing 5% β‐mercaptoethanol.

    Article Title: Bleomycin hydrolase regulates the release of chemokines important for inflammation and wound healing by keratinocytes
    Article Snippet: .. Western blot For Western blot analysis of protein expression, 30 ug of total protein lysates was mixed with NuPAGE LDS Sample buffer (Invitrogen) and NuPAGE Sample Reducing Agent (Invitrogen) and heated for 10 minutes, 70 °C. .. The samples were loaded onto NuPAGE 4–12% Bis-Tris Protein Gels (Invitrogen) and run with NuPAGE MOPS SDS Running buffer (Invitrogen) according to the NuPAGE Novex electrophoresis program.

    Article Title: Autophagy and mitochondrial biogenesis impairment contribute to age-dependent liver injury in experimental sepsis: dysregulation of AMP-activated protein kinase pathway
    Article Snippet: Paragraph title: Western blot analysis ... Cytosol and nuclear extracts were boiled in equal volumes of NuPAGE LDS Sample Buffer (Thermo Fisher Scientific) and 25–40 μg of protein loaded per lane on a 16% Tris-glycine gradient gel.

    Article Title: Protein kinase C epsilon contributes to basal and sensitizing responses of TRPV1 to capsaicin in rat dorsal root ganglion neurons
    Article Snippet: Paragraph title: Western blotting ... Vector-infected U2OS cells (MOI 3) were harvested at 16 h post-infection (hpi) with 1× NuPAGE LDS Sample Buffer in PBS (Invitrogen, Carlsbad, CA, USA) and heated at 70°C for 10 min.

    Article Title: Efficacy of Multi-exon Skipping Treatment in Duchenne Muscular Dystrophy Dog Model Neonates
    Article Snippet: .. Protein was prepared by adding NuPAGE LDS Sample Buffer (Thermo Fisher) and NuPAGE Sample Reducing Agent (Thermo Fisher) at 1× final concentrations into the samples and then incubating at 70°C for 10 min. Western blotting was performed as described previously. ..

    Protease Inhibitor:

    Article Title: The AKT isoforms 1 and 2 drive B cell fate decisions during the germinal center response
    Article Snippet: For analysis of protein expression in whole cells, whole cell lysates were prepared by direct lysing with radio immunoprecipitation assay buffer plus complete protease inhibitor mixture (Roche). .. The samples were boiled in NuPAGE LDS Sample Buffer (Invitrogen) supplemented with β-mercaptoethanol and then were separated by 4–12% polyacrylamide Bis-Tris gel (Bio-Rad) and transferred onto polyvinylidene difluoride membrane (EMD Millipore).

    Article Title: Lentiviral Vector Production Titer Is Not Limited in HEK293T by Induced Intracellular Innate Immunity
    Article Snippet: Immunoblots Cell extracts were prepared by washing 5 × 106 cells with cold PBS and resuspending cell pellets in radioimmunoprecipitation assay (RIPA) buffer supplemented with Halt protease inhibitor cocktail (Thermo Fisher Scientific). .. Whole cell lysis was carried out on ice for 15 min and lysates cleared by centrifugation in a tabletop centrifuge at full speed for 5 min. Total protein was quantified using BCA Protein Assay Reagents (Pierce Thermo Fisher Scientific) and 25 μg total protein extract was heated at 95°C for 5 min with NuPAGE LDS Sample Buffer (Thermo Fisher Scientific) and then loaded on a Novex NuPAGE 4%–12% Bis-Tris gel (Thermo Fisher Scientific).

    Article Title: TGFβ1‐induced cell motility but not cell proliferation is mediated through Cten in colorectal cancer
    Article Snippet: Cell lysates were prepared using RIPA lysis buffer (Thermo Fisher Scientific) supplemented with phosphatase and protease inhibitor (Thermo Fisher Scientific). .. Fifty micrograms of protein was added to NUPAGE LDS Sample Buffer (Thermo Fisher Scientific) containing 5% β‐mercaptoethanol.

    Article Title: TGFβ1‐induced cell motility but not cell proliferation is mediated through Cten in colorectal cancer
    Article Snippet: Cell lysates were prepared using RIPA lysis buffer (Thermo Fisher Scientific) supplemented with phosphatase and protease inhibitor (Thermo Fisher Scientific). .. Fifty micrograms of protein was added to NUPAGE LDS Sample Buffer (Thermo Fisher Scientific) containing 5% β‐mercaptoethanol.

    Article Title: CAT tails drive degradation of stalled polypeptides on and off the ribosome
    Article Snippet: The resulting “grindate” was re-solubilized in TEV buffer (25mM HEPES-KOH pH 7.4, 150mM KOAc, 0.5mM EDTA, 10% glycerol, 0.04% Antifoam-B, and EDTA-Free Pierce Protease Inhibitor Mini Tablets; added 1:1 vol:mass) to produce lysate. .. 4x NuPage LDS Sample Buffer (Thermo Fisher Scientific) with 5% β-mercaptoethanol was added at 1:1 vol:vol before boiling at 95°C for 5 min to denature the samples.

    other:

    Article Title: Mapping Exosome-Substrate Interactions In Vivo by UV Cross-Linking.
    Article Snippet: The RNA exosome complex functions in both the accurate processing and rapid degradation of many classes of RNA in eukaryotes and Archaea.

    Protein Concentration:

    Article Title: The AKT isoforms 1 and 2 drive B cell fate decisions during the germinal center response
    Article Snippet: Protein concentration of subcellular fractionation and whole cell lysates was quantitated using the BCA protein assay kit (Pierce). .. The samples were boiled in NuPAGE LDS Sample Buffer (Invitrogen) supplemented with β-mercaptoethanol and then were separated by 4–12% polyacrylamide Bis-Tris gel (Bio-Rad) and transferred onto polyvinylidene difluoride membrane (EMD Millipore).

    Sonication:

    Article Title: p53 controls genomic stability and temporal differentiation of human neural stem cells and affects neural organization in human brain organoids
    Article Snippet: .. Western blot Whole-cell lysates were prepared by lysing cell pellets in 1×NuPAGE LDS Sample buffer including protease and phosphatase inhibitor (ThermoFisher Scientific, 78440), incubated 5 min at 90 °C and 3 rounds of sonication for 5 min. Total protein was loaded on a 4–12% gradient Bis/Tris gel (ThermoFisher Scientific, NP0322BOX). .. Proteins were transferred to a nitrocellulose membrane, using the Trans-Blot Turbo Transfer System (Bio-Rad, 1704150).

    Nucleic Acid Electrophoresis:

    Article Title: TGFβ1‐induced cell motility but not cell proliferation is mediated through Cten in colorectal cancer
    Article Snippet: Fifty micrograms of protein was added to NUPAGE LDS Sample Buffer (Thermo Fisher Scientific) containing 5% β‐mercaptoethanol. .. Following this, the protein samples were fractionated on a precast 4%‐12% NUPAGE Bis‐Tris‐HCl buffered (pH 6.4) polyacrylamide gel (Thermo Fisher Scientific) using the NUPAGE gel electrophoresis system with NUPAGE MOPS SDS Running Buffer (Thermo Fisher Scientific, Carlsbad, CA) at 125 V for 90 minutes.

    Article Title: TGFβ1‐induced cell motility but not cell proliferation is mediated through Cten in colorectal cancer
    Article Snippet: Fifty micrograms of protein was added to NUPAGE LDS Sample Buffer (Thermo Fisher Scientific) containing 5% β‐mercaptoethanol. .. Following this, the protein samples were fractionated on a precast 4%‐12% NUPAGE Bis‐Tris‐HCl buffered (pH 6.4) polyacrylamide gel (Thermo Fisher Scientific) using the NUPAGE gel electrophoresis system with NUPAGE MOPS SDS Running Buffer (Thermo Fisher Scientific, Carlsbad, CA) at 125 V for 90 minutes.

    Isolation:

    Article Title: The long noncoding RNA NEAT1 and nuclear paraspeckles are up-regulated by the transcription factor HSF1 in the heat shock response
    Article Snippet: Whole-cell extracts (WCE) were made by lysing cells directly in 2× NuPAGE LDS Sample Buffer (ThermoFisher Scientific). .. Nuclear extracts (NE) were isolated using the NE-PERTM nuclear and cytoplasmic extraction kit (ThermoFisher Scientific) according to the manufacturer's instruction.

    Protein Extraction:

    Article Title: Efficacy of Multi-exon Skipping Treatment in Duchenne Muscular Dystrophy Dog Model Neonates
    Article Snippet: For protein extraction, 100 μL of a high SDS lysis buffer (10% SDS, 70 mM Tris-HCl [at pH 6.7], 5 mM EDTA at pH 8.0, 5% β-mercaptoethanol in water) with proteinase inhibitor cocktail (Roche) was added to 20-μm frozen muscle sections. .. Protein was prepared by adding NuPAGE LDS Sample Buffer (Thermo Fisher) and NuPAGE Sample Reducing Agent (Thermo Fisher) at 1× final concentrations into the samples and then incubating at 70°C for 10 min. Western blotting was performed as described previously.

    Article Title: Efficacy of Multi-exon Skipping Treatment in Duchenne Muscular Dystrophy Dog Model Neonates
    Article Snippet: For protein extraction, 100 μL of a high SDS lysis buffer (10% SDS, 70 mM Tris-HCl [at pH 6.7], 5 mM EDTA at pH 8.0, 5% β-mercaptoethanol in water) with proteinase inhibitor cocktail (Roche) was added to 20-μm frozen muscle sections. .. Protein was prepared by adding NuPAGE LDS Sample Buffer (Thermo Fisher) and NuPAGE Sample Reducing Agent (Thermo Fisher) at 1× final concentrations into the samples and then incubating at 70°C for 10 min. Western blotting was performed as described previously.

    Staining:

    Article Title: The long noncoding RNA neuroLNC regulates presynaptic activity by interacting with the neurodegeneration-associated protein TDP-43
    Article Snippet: MS identification of protein interactors following ChIRP/RNA interactome analysis For proteomic analysis, we initially reversed the cross-linking by boiling the beads in a final concentration of 1× NuPAGE LDS Sample Buffer (Thermo Fisher Scientific) at 95°C for 30 min on a thermomixer (at 750 rpm). .. The gels were stained overnight with Coomassie G250, and for each lane, five to six pieces were cut.

    Plasmid Preparation:

    Article Title: Protein kinase C epsilon contributes to basal and sensitizing responses of TRPV1 to capsaicin in rat dorsal root ganglion neurons
    Article Snippet: .. Vector-infected U2OS cells (MOI 3) were harvested at 16 h post-infection (hpi) with 1× NuPAGE LDS Sample Buffer in PBS (Invitrogen, Carlsbad, CA, USA) and heated at 70°C for 10 min. ..

    Radio Immunoprecipitation:

    Article Title: The AKT isoforms 1 and 2 drive B cell fate decisions during the germinal center response
    Article Snippet: For analysis of protein expression in whole cells, whole cell lysates were prepared by direct lysing with radio immunoprecipitation assay buffer plus complete protease inhibitor mixture (Roche). .. The samples were boiled in NuPAGE LDS Sample Buffer (Invitrogen) supplemented with β-mercaptoethanol and then were separated by 4–12% polyacrylamide Bis-Tris gel (Bio-Rad) and transferred onto polyvinylidene difluoride membrane (EMD Millipore).

    Article Title: Lentiviral Vector Production Titer Is Not Limited in HEK293T by Induced Intracellular Innate Immunity
    Article Snippet: Immunoblots Cell extracts were prepared by washing 5 × 106 cells with cold PBS and resuspending cell pellets in radioimmunoprecipitation assay (RIPA) buffer supplemented with Halt protease inhibitor cocktail (Thermo Fisher Scientific). .. Whole cell lysis was carried out on ice for 15 min and lysates cleared by centrifugation in a tabletop centrifuge at full speed for 5 min. Total protein was quantified using BCA Protein Assay Reagents (Pierce Thermo Fisher Scientific) and 25 μg total protein extract was heated at 95°C for 5 min with NuPAGE LDS Sample Buffer (Thermo Fisher Scientific) and then loaded on a Novex NuPAGE 4%–12% Bis-Tris gel (Thermo Fisher Scientific).

    Concentration Assay:

    Article Title: The long noncoding RNA neuroLNC regulates presynaptic activity by interacting with the neurodegeneration-associated protein TDP-43
    Article Snippet: .. MS identification of protein interactors following ChIRP/RNA interactome analysis For proteomic analysis, we initially reversed the cross-linking by boiling the beads in a final concentration of 1× NuPAGE LDS Sample Buffer (Thermo Fisher Scientific) at 95°C for 30 min on a thermomixer (at 750 rpm). .. After boiling, the samples were loaded on precast NuPAGE 4 to 12% Bis-Tris Protein Gels (Thermo Fisher Scientific) and separated for 20 min at constant voltage (200 V).

    Fractionation:

    Article Title: The AKT isoforms 1 and 2 drive B cell fate decisions during the germinal center response
    Article Snippet: Paragraph title: Subcellular fractionation and Western blot analysis ... The samples were boiled in NuPAGE LDS Sample Buffer (Invitrogen) supplemented with β-mercaptoethanol and then were separated by 4–12% polyacrylamide Bis-Tris gel (Bio-Rad) and transferred onto polyvinylidene difluoride membrane (EMD Millipore).

    Lysis:

    Article Title: Lentiviral Vector Production Titer Is Not Limited in HEK293T by Induced Intracellular Innate Immunity
    Article Snippet: .. Whole cell lysis was carried out on ice for 15 min and lysates cleared by centrifugation in a tabletop centrifuge at full speed for 5 min. Total protein was quantified using BCA Protein Assay Reagents (Pierce Thermo Fisher Scientific) and 25 μg total protein extract was heated at 95°C for 5 min with NuPAGE LDS Sample Buffer (Thermo Fisher Scientific) and then loaded on a Novex NuPAGE 4%–12% Bis-Tris gel (Thermo Fisher Scientific). .. Wet transfer was performed using the X-Cell SureLock Blot module (Thermo Fisher Scientific) onto Immun-blot PVDF Membrane (BioRad, Hercules, CA, USA).

    Article Title: Efficacy of Multi-exon Skipping Treatment in Duchenne Muscular Dystrophy Dog Model Neonates
    Article Snippet: For protein extraction, 100 μL of a high SDS lysis buffer (10% SDS, 70 mM Tris-HCl [at pH 6.7], 5 mM EDTA at pH 8.0, 5% β-mercaptoethanol in water) with proteinase inhibitor cocktail (Roche) was added to 20-μm frozen muscle sections. .. Protein was prepared by adding NuPAGE LDS Sample Buffer (Thermo Fisher) and NuPAGE Sample Reducing Agent (Thermo Fisher) at 1× final concentrations into the samples and then incubating at 70°C for 10 min. Western blotting was performed as described previously.

    Article Title: TGFβ1‐induced cell motility but not cell proliferation is mediated through Cten in colorectal cancer
    Article Snippet: Cell lysates were prepared using RIPA lysis buffer (Thermo Fisher Scientific) supplemented with phosphatase and protease inhibitor (Thermo Fisher Scientific). .. Fifty micrograms of protein was added to NUPAGE LDS Sample Buffer (Thermo Fisher Scientific) containing 5% β‐mercaptoethanol.

    Article Title: TGFβ1‐induced cell motility but not cell proliferation is mediated through Cten in colorectal cancer
    Article Snippet: Cell lysates were prepared using RIPA lysis buffer (Thermo Fisher Scientific) supplemented with phosphatase and protease inhibitor (Thermo Fisher Scientific). .. Fifty micrograms of protein was added to NUPAGE LDS Sample Buffer (Thermo Fisher Scientific) containing 5% β‐mercaptoethanol.

    Article Title: Efficacy of Multi-exon Skipping Treatment in Duchenne Muscular Dystrophy Dog Model Neonates
    Article Snippet: For protein extraction, 100 μL of a high SDS lysis buffer (10% SDS, 70 mM Tris-HCl [at pH 6.7], 5 mM EDTA at pH 8.0, 5% β-mercaptoethanol in water) with proteinase inhibitor cocktail (Roche) was added to 20-μm frozen muscle sections. .. Protein was prepared by adding NuPAGE LDS Sample Buffer (Thermo Fisher) and NuPAGE Sample Reducing Agent (Thermo Fisher) at 1× final concentrations into the samples and then incubating at 70°C for 10 min. Western blotting was performed as described previously.

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    Thermo Fisher lithium dodecyl sulfate lds
    Stoichiometry of histone acetylation. a The diagram shows all histone acetylation sites whose stoichiometry was determined in this study. The sites are ordered by descending stoichiometry. Note that high stoichiometry sites occur on the N-termini of core histones. b The stoichiometry of histone acetylation sites as determined in four independent studies 17 – 20 . c An anti-acetylated lysine immunoblot of <t>HeLa</t> whole cell lysate. Cells were boiled in 2% <t>LDS</t> to ensure histone extraction. Histones are annotated based on their expected molecular weight. d Histone acetylation sites constitute a majority of acetylated lysine residues in cells. Stoichiometry and protein copy numbers were used to calculate the number of acetylated lysine residues for the indicated classes of proteins. Source data are provided as a Source Data file
    Lithium Dodecyl Sulfate Lds, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 834 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/lithium dodecyl sulfate lds/product/Thermo Fisher
    Average 99 stars, based on 834 article reviews
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    lithium dodecyl sulfate lds - by Bioz Stars, 2020-03
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    Stoichiometry of histone acetylation. a The diagram shows all histone acetylation sites whose stoichiometry was determined in this study. The sites are ordered by descending stoichiometry. Note that high stoichiometry sites occur on the N-termini of core histones. b The stoichiometry of histone acetylation sites as determined in four independent studies 17 – 20 . c An anti-acetylated lysine immunoblot of HeLa whole cell lysate. Cells were boiled in 2% LDS to ensure histone extraction. Histones are annotated based on their expected molecular weight. d Histone acetylation sites constitute a majority of acetylated lysine residues in cells. Stoichiometry and protein copy numbers were used to calculate the number of acetylated lysine residues for the indicated classes of proteins. Source data are provided as a Source Data file

    Journal: Nature Communications

    Article Title: Analysis of human acetylation stoichiometry defines mechanistic constraints on protein regulation

    doi: 10.1038/s41467-019-09024-0

    Figure Lengend Snippet: Stoichiometry of histone acetylation. a The diagram shows all histone acetylation sites whose stoichiometry was determined in this study. The sites are ordered by descending stoichiometry. Note that high stoichiometry sites occur on the N-termini of core histones. b The stoichiometry of histone acetylation sites as determined in four independent studies 17 – 20 . c An anti-acetylated lysine immunoblot of HeLa whole cell lysate. Cells were boiled in 2% LDS to ensure histone extraction. Histones are annotated based on their expected molecular weight. d Histone acetylation sites constitute a majority of acetylated lysine residues in cells. Stoichiometry and protein copy numbers were used to calculate the number of acetylated lysine residues for the indicated classes of proteins. Source data are provided as a Source Data file

    Article Snippet: Histone western blot HeLa cells were lysed by boiling in 2% lithium dodecyl sulfate (LDS) (1x NuPAGE LDS Sample Buffer, ThermoFischer Scientific), followed by sonication to disrupt genomic DNA.

    Techniques: Molecular Weight