pierce ecl western blotting substrate  (Thermo Fisher)


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    Structured Review

    Thermo Fisher pierce ecl western blotting substrate
    Pierce Ecl Western Blotting Substrate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1193 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pierce ecl western blotting substrate/product/Thermo Fisher
    Average 99 stars, based on 1193 article reviews
    Price from $9.99 to $1999.99
    pierce ecl western blotting substrate - by Bioz Stars, 2020-01
    99/100 stars

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    Centrifugation:

    Article Title: Akap1 genetic deletion increases the severity of hyperoxia-induced acute lung injury in mice
    Article Snippet: Frozen lung tissue was pulverized and lysed in protein extraction buffer (20 mM Tris·HCl, pH 7.4, 150 mM NaCl, and 0.5% Triton X-100), and the supernatant, containing soluble protein, was collected by high-speed centrifugation (14,000 g ) for 15 min at 4°C. .. The proteins were visualized using Pierce ECL Western blotting substrate (Thermo Fisher Scientifics).

    Stable Transfection:

    Article Title: Tumor suppression of novel anti–PD-1 antibodies mediated through CD28 costimulatory pathway
    Article Snippet: For each condition, 10 million Jurkat cells stably expressing PD-1 (BPS Bioscience) were either left unstimulated or stimulated with biotinylated anti-CD3 (5 µg/ml; OKT3; eBioscience) and anti-CD28 (5 µg/ml; CD28.2; eBioscience) then aggregated with the addition of 50 µg/ml of avidin (Invitrogen). .. Immunoblotting was performed by staining with the primary Ab overnight at 4°C followed by washing steps and then incubation with the HRP-conjugated secondary Ab and detection with the Pierce ECL Western blotting substrate (Thermo Fisher Scientific) in accordance with the manufacturer’s protocol.

    Blocking Assay:

    Article Title: TLR4 (toll-like receptor 4) activation suppresses autophagy through inhibition of FOXO3 and impairs phagocytic capacity of microglia
    Article Snippet: The membranes were then incubated with primary antibodies overnight at 4°C in a shaking incubator, washed with TBST 3 times (10 min each) and incubated for 1 h at room temperature with peroxidase-conjugated secondary antibodies diluted in blocking solution. .. After washing, proteins of interest were detected using either Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, 32106) or WesternBright ECL (Advansta, K-12045-D50).

    Electrophoresis:

    Article Title: TLR4 (toll-like receptor 4) activation suppresses autophagy through inhibition of FOXO3 and impairs phagocytic capacity of microglia
    Article Snippet: Samples were separated by SDS gel electrophoresis and electro-transferred onto polyvinylidene difluoride membrane in a semi-dry electrophoretic transfer cell (Bio-Rad). .. After washing, proteins of interest were detected using either Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, 32106) or WesternBright ECL (Advansta, K-12045-D50).

    Article Title: Comparative flavivirus-host protein interaction mapping reveals mechanisms of dengue and Zika virus pathogenesis
    Article Snippet: Following electrophoresis, proteins were transferred to PVDF (BioRad) with 20% methanol transfer buffer. .. Signal was detected using Pierce ECL Western Blotting Substrate (Thermo Fisher).

    Article Title: Decellularized Extracellular Matrix Hydrogels as a Delivery Platform for MicroRNA and Extracellular Vesicle Therapeutics
    Article Snippet: Western blot was performed using the XCell SureLock Mini-Cell Electrophoresis System filled with 1X NuPAGE Transfer Buffer (Thermo Fisher Scientific) in 10% methanol in deionized water. .. Prior to imaging, the membrane was incubated with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific) for 1 minute and imaged with the Bio-Rad ChemiDoc MP System using Image Lab 3.0 software.

    Incubation:

    Article Title: Akap1 genetic deletion increases the severity of hyperoxia-induced acute lung injury in mice
    Article Snippet: Membranes were blocked in 5% skim milk with TBS-T followed by incubation with appropriate primary and secondary antibodies. .. The proteins were visualized using Pierce ECL Western blotting substrate (Thermo Fisher Scientifics).

    Article Title: TLR4 (toll-like receptor 4) activation suppresses autophagy through inhibition of FOXO3 and impairs phagocytic capacity of microglia
    Article Snippet: The membranes were then incubated with primary antibodies overnight at 4°C in a shaking incubator, washed with TBST 3 times (10 min each) and incubated for 1 h at room temperature with peroxidase-conjugated secondary antibodies diluted in blocking solution. .. After washing, proteins of interest were detected using either Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, 32106) or WesternBright ECL (Advansta, K-12045-D50).

    Article Title: Tumor suppression of novel anti–PD-1 antibodies mediated through CD28 costimulatory pathway
    Article Snippet: .. Immunoblotting was performed by staining with the primary Ab overnight at 4°C followed by washing steps and then incubation with the HRP-conjugated secondary Ab and detection with the Pierce ECL Western blotting substrate (Thermo Fisher Scientific) in accordance with the manufacturer’s protocol. .. Blots were imaged with the Fusion FX Vilber Lurmat camera (Witec), and the density of individual bands was evaluated using ImageJ software.

    Article Title: Comparative flavivirus-host protein interaction mapping reveals mechanisms of dengue and Zika virus pathogenesis
    Article Snippet: Secondary HRP (BioRad) incubation occurred 1:10000 dilution for one hour at room temperature. .. Signal was detected using Pierce ECL Western Blotting Substrate (Thermo Fisher).

    Article Title: Ponatinib shows potent antitumor activity in small cell carcinoma of the ovary hypercalcemic type (SCCOHT) through multi-kinase inhibition
    Article Snippet: Three hundred μg of lysate was incubated on each blot overnight, and bound phosphorylated protein was detected using HRP-conjugated anti-phospho-tyrosine antibody. .. Arrays were developed using Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific), and images were quantified using ImageJ software (NIH) using built-in gel quantification and background subtraction tools.

    Article Title: Accelerated decline in cognition in a mouse model of increased oxidative stress
    Article Snippet: .. This was followed by incubation with anti-rabbit IgG HRP conjugated antibody and the blot was developed using Pierce ECL Western Blotting Substrate (ThermoFisher Scientific, Waltham, MA). .. Images were taken using a G:BOX imaging system (Syngene, Frederick, MD) and quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD, USA).

    Article Title: Decellularized Extracellular Matrix Hydrogels as a Delivery Platform for MicroRNA and Extracellular Vesicle Therapeutics
    Article Snippet: .. Prior to imaging, the membrane was incubated with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific) for 1 minute and imaged with the Bio-Rad ChemiDoc MP System using Image Lab 3.0 software. .. A 96-well plate was coated with 0.1% porcine gelatin and 7500 hCPCs per well were seeded and incubated in growth media overnight.

    Article Title: Mitochondrial Uncoupling Protein 2 Knock-out Promotes Mitophagy to Decrease Retinal Ganglion Cell Death in a Mouse Model of Glaucoma
    Article Snippet: .. Membranes were washed in TBS-T and incubated in secondary antibody for 2 h at room temperature before generation of chemiluminescent bands with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, catalog #32106) and visualization on photosensitive films (Thermo Fisher Scientific, catalog #34091). ..

    Article Title: SIV-Induced Immune Activation and Metabolic Alterations in the Dorsal Root Ganglia During Acute Infection
    Article Snippet: Membranes were then incubated with primary against glutaminase ([GLS], rabbit monoclonal; Abcam, Cambridge, UK), glutamine synthetase ([GS], rabbit polyclonal, Abcam), superoxide dismutase-2 ([SOD2], rabbit polyclonal; Abcam), βIII tubulin (clone 5G8; Promega), and β-actin (clone AC-15; Sigma), followed by horseradish peroxidase-conjugated secondary antibodies (goat anti-rabbit IgG and goat anti-mouse IgG; Dako, Carpinteria, CA). .. Membranes were then washed in TBST and developed with Pierce ECL Western Blotting Substrate (Thermo Fisher) prior to exposure to radiography film.

    Article Title: Anti‐inflammatory effects of miR‐21 in the macrophage response to peritonitis
    Article Snippet: Following incubation with each antibody, membranes were washed and subsequently incubated with rabbit anti‐mouse primary antibody for housekeeping protein β‐actin (1:10,000) or GAPDH (1:10,000) and then with goat anti‐rabbit secondary antibody (1:5,000). .. Pierce ECL Western blotting substrate (Thermo Fisher) was applied to the membrane as per the manufacturer's instructions.

    Expressing:

    Article Title: Tumor suppression of novel anti–PD-1 antibodies mediated through CD28 costimulatory pathway
    Article Snippet: For each condition, 10 million Jurkat cells stably expressing PD-1 (BPS Bioscience) were either left unstimulated or stimulated with biotinylated anti-CD3 (5 µg/ml; OKT3; eBioscience) and anti-CD28 (5 µg/ml; CD28.2; eBioscience) then aggregated with the addition of 50 µg/ml of avidin (Invitrogen). .. Immunoblotting was performed by staining with the primary Ab overnight at 4°C followed by washing steps and then incubation with the HRP-conjugated secondary Ab and detection with the Pierce ECL Western blotting substrate (Thermo Fisher Scientific) in accordance with the manufacturer’s protocol.

    Article Title: Comparative flavivirus-host protein interaction mapping reveals mechanisms of dengue and Zika virus pathogenesis
    Article Snippet: C-terminally Strep II-tagged flavivirus NS5 constructs were expressed in HEK 293T and affinity purified as described for viral proteins in Expression and affinity purification of viral proteins . .. Signal was detected using Pierce ECL Western Blotting Substrate (Thermo Fisher).

    Article Title: Decellularized Extracellular Matrix Hydrogels as a Delivery Platform for MicroRNA and Extracellular Vesicle Therapeutics
    Article Snippet: Paragraph title: EV Bioactivity – Stimulation of pERK Expression: ... Prior to imaging, the membrane was incubated with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific) for 1 minute and imaged with the Bio-Rad ChemiDoc MP System using Image Lab 3.0 software.

    Article Title: SIV-Induced Immune Activation and Metabolic Alterations in the Dorsal Root Ganglia During Acute Infection
    Article Snippet: Membranes were then washed in TBST and developed with Pierce ECL Western Blotting Substrate (Thermo Fisher) prior to exposure to radiography film. .. To account for blot-to-blot variation, a standard DRG lysate was included in each blot for normalization and β-actin expression was utilized as an in-lane loading control.

    BIA-KA:

    Article Title: Akap1 genetic deletion increases the severity of hyperoxia-induced acute lung injury in mice
    Article Snippet: Protein was estimated by BCA Protein Assay kit (Pierce, Rockford, IL). .. The proteins were visualized using Pierce ECL Western blotting substrate (Thermo Fisher Scientifics).

    Article Title: TLR4 (toll-like receptor 4) activation suppresses autophagy through inhibition of FOXO3 and impairs phagocytic capacity of microglia
    Article Snippet: BCA protein assay reagents (Thermo Fisher Scientific, 23225) were used to measure protein concentration. .. After washing, proteins of interest were detected using either Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, 32106) or WesternBright ECL (Advansta, K-12045-D50).

    Modification:

    Article Title: Accelerated decline in cognition in a mouse model of increased oxidative stress
    Article Snippet: For the detection of 4-HNE modified proteins, equal amounts of protein (40 μg/lane) were separated by SDS–PAGE, transferred to polyvinylidene difluoride membranes and treated with 250 mM sodium borohydride in 100 mM (3-(N-morpholino)propanesulfonic acid, MOPS), pH 8.0 for 15 min. .. This was followed by incubation with anti-rabbit IgG HRP conjugated antibody and the blot was developed using Pierce ECL Western Blotting Substrate (ThermoFisher Scientific, Waltham, MA).

    Western Blot:

    Article Title: Akap1 genetic deletion increases the severity of hyperoxia-induced acute lung injury in mice
    Article Snippet: .. The proteins were visualized using Pierce ECL Western blotting substrate (Thermo Fisher Scientifics). .. Protein bands were scanned, and densitometry was analyzed with NIH ImageJ software.

    Article Title: TLR4 (toll-like receptor 4) activation suppresses autophagy through inhibition of FOXO3 and impairs phagocytic capacity of microglia
    Article Snippet: .. After washing, proteins of interest were detected using either Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, 32106) or WesternBright ECL (Advansta, K-12045-D50). .. Cells were lysed using the QIAzol Lysis Reagent (Qiagen, 79306), and RNA was isolated following the manufacturer’s instructions. cDNA was synthesized using the ImProm-II Reverse Transcriptase kit (Promega, A3802) and oligo-dT primers.

    Article Title: Tumor suppression of novel anti–PD-1 antibodies mediated through CD28 costimulatory pathway
    Article Snippet: .. Immunoblotting was performed by staining with the primary Ab overnight at 4°C followed by washing steps and then incubation with the HRP-conjugated secondary Ab and detection with the Pierce ECL Western blotting substrate (Thermo Fisher Scientific) in accordance with the manufacturer’s protocol. .. Blots were imaged with the Fusion FX Vilber Lurmat camera (Witec), and the density of individual bands was evaluated using ImageJ software.

    Article Title: Ripor2 is involved in auditory hair cell stereociliary bundle structure and orientation
    Article Snippet: .. The results were visualized by chemiluminescence using Pierce ECL Western Blotting Substrate (Thermo Scientific). .. For western blots assessing abundance of endogenous cochlear proteins we followed the protocol as previously described but using the appropriate primary antibodies as corresponded.

    Article Title: Comparative flavivirus-host protein interaction mapping reveals mechanisms of dengue and Zika virus pathogenesis
    Article Snippet: .. Signal was detected using Pierce ECL Western Blotting Substrate (Thermo Fisher). .. For siRNA knockdown studies, U2OS cells were seeded at 5000 cells/well in 50 μL media per well in 384-well for siRNA knockdown. siRNAs (Sigma) ( ) were transfected with HiPerFect (Qiagen) according to manufacturer recommendations at a final siRNA concentration of 20 nM.

    Article Title: Ponatinib shows potent antitumor activity in small cell carcinoma of the ovary hypercalcemic type (SCCOHT) through multi-kinase inhibition
    Article Snippet: .. Arrays were developed using Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific), and images were quantified using ImageJ software (NIH) using built-in gel quantification and background subtraction tools. ..

    Article Title: Effects and Mechanism of Nicotinamide Against UVA- and/or UVB-mediated DNA Damages in Normal Melanocytes
    Article Snippet: .. Proteins were detected by chemiluminescence using Pierce ECL Western Blotting Substrate, or SuperSignal West Femto Chemiluminescent Substrates (Pierce, Rockford, IL). .. The quantification of the Western blot bands was performed using ImageJ (US National Institutes of Health, Bethesda, MD, USA).

    Article Title: Accelerated decline in cognition in a mouse model of increased oxidative stress
    Article Snippet: .. This was followed by incubation with anti-rabbit IgG HRP conjugated antibody and the blot was developed using Pierce ECL Western Blotting Substrate (ThermoFisher Scientific, Waltham, MA). .. Images were taken using a G:BOX imaging system (Syngene, Frederick, MD) and quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD, USA).

    Article Title: Envelope Protein Glycosylation Mediates Zika Virus Pathogenesis
    Article Snippet: .. Pierce ECL Western blotting substrate was used to detect HRP (Thermo). .. Western and lectin blots were imaged on a ChemiDoc XRS+ system (Bio-Rad).

    Article Title: Live-cell imaging of single mRNA dynamics using split superfolder green fluorescent proteins with minimal background
    Article Snippet: .. Pierce ECL western blotting substrate (Thermo Scientific) was used for HRP detection. .. The western blots were imaged by a LAS 4000 (GE Healthcare Life Sciences).

    Article Title: Decellularized Extracellular Matrix Hydrogels as a Delivery Platform for MicroRNA and Extracellular Vesicle Therapeutics
    Article Snippet: .. Prior to imaging, the membrane was incubated with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific) for 1 minute and imaged with the Bio-Rad ChemiDoc MP System using Image Lab 3.0 software. .. A 96-well plate was coated with 0.1% porcine gelatin and 7500 hCPCs per well were seeded and incubated in growth media overnight.

    Article Title: Mitochondrial Uncoupling Protein 2 Knock-out Promotes Mitophagy to Decrease Retinal Ganglion Cell Death in a Mouse Model of Glaucoma
    Article Snippet: .. Membranes were washed in TBS-T and incubated in secondary antibody for 2 h at room temperature before generation of chemiluminescent bands with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, catalog #32106) and visualization on photosensitive films (Thermo Fisher Scientific, catalog #34091). ..

    Article Title: SIV-Induced Immune Activation and Metabolic Alterations in the Dorsal Root Ganglia During Acute Infection
    Article Snippet: .. Membranes were then washed in TBST and developed with Pierce ECL Western Blotting Substrate (Thermo Fisher) prior to exposure to radiography film. .. Because GS, βIII tubulin, and β-actin have similar molecular weights, the membranes were first probed for GS, stripped using Restore PLUS Western Blot Stripping Buffer (Thermo Fisher) for 15 minutes at room temperature, then re-probed for βIII-tubulin, followed by β-actin.

    Article Title: Inactivation of mTORC2 in macrophages is a signature of colorectal cancer that promotes tumorigenesis
    Article Snippet: .. Proteins were detected by chemiluminescence using the Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific). .. The reaction was visualized using Amersham Hyperfilm ECL (GE Healthcare) and the Medical X-ray Processor 2000 (Kodak).

    Article Title: Anti‐inflammatory effects of miR‐21 in the macrophage response to peritonitis
    Article Snippet: .. Pierce ECL Western blotting substrate (Thermo Fisher) was applied to the membrane as per the manufacturer's instructions. .. CL‐XPosure Film (Thermo Fisher) was used to expose the membrane for 15 min.

    Derivative Assay:

    Article Title: Accelerated decline in cognition in a mouse model of increased oxidative stress
    Article Snippet: The antibody recognizes cysteine, lysine, and histidine 4-HNE protein adducts and is highly specific to 4-HNE derived protein adducts (gift from Dr. Luke Szweda, Oklahoma Medical Research Foundation) (Uchida et al. ). .. This was followed by incubation with anti-rabbit IgG HRP conjugated antibody and the blot was developed using Pierce ECL Western Blotting Substrate (ThermoFisher Scientific, Waltham, MA).

    Electrotransfer:

    Article Title: Akap1 genetic deletion increases the severity of hyperoxia-induced acute lung injury in mice
    Article Snippet: Equal amounts of protein (20 µg) were subjected SDS-PAGE by using 4–20% Tris-glycine gels (Bio-Rad), followed by electrotransfer to PVDF membrane. .. The proteins were visualized using Pierce ECL Western blotting substrate (Thermo Fisher Scientifics).

    Protease Inhibitor:

    Article Title: Tumor suppression of novel anti–PD-1 antibodies mediated through CD28 costimulatory pathway
    Article Snippet: Cell membranes where then lysed with ice cold PBS containing 1% Triton X-100, protease inhibitor cocktail (Complete; Roche) and a phosphatase inhibitor cocktail followed by a 15-min incubation on ice with frequent mixing. .. Immunoblotting was performed by staining with the primary Ab overnight at 4°C followed by washing steps and then incubation with the HRP-conjugated secondary Ab and detection with the Pierce ECL Western blotting substrate (Thermo Fisher Scientific) in accordance with the manufacturer’s protocol.

    Article Title: Mitochondrial Uncoupling Protein 2 Knock-out Promotes Mitophagy to Decrease Retinal Ganglion Cell Death in a Mouse Model of Glaucoma
    Article Snippet: Briefly, dissected retinal tissue was homogenized in RIPA buffer supplemented with Halt protease inhibitor mixture (Thermo Fisher Scientific, catalog #78430) and a phosphatase inhibitor mixture set II (EMD Millipore, catalog #524625). .. Membranes were washed in TBS-T and incubated in secondary antibody for 2 h at room temperature before generation of chemiluminescent bands with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, catalog #32106) and visualization on photosensitive films (Thermo Fisher Scientific, catalog #34091).

    Imaging:

    Article Title: Accelerated decline in cognition in a mouse model of increased oxidative stress
    Article Snippet: This was followed by incubation with anti-rabbit IgG HRP conjugated antibody and the blot was developed using Pierce ECL Western Blotting Substrate (ThermoFisher Scientific, Waltham, MA). .. Images were taken using a G:BOX imaging system (Syngene, Frederick, MD) and quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD, USA).

    Article Title: Decellularized Extracellular Matrix Hydrogels as a Delivery Platform for MicroRNA and Extracellular Vesicle Therapeutics
    Article Snippet: .. Prior to imaging, the membrane was incubated with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific) for 1 minute and imaged with the Bio-Rad ChemiDoc MP System using Image Lab 3.0 software. .. A 96-well plate was coated with 0.1% porcine gelatin and 7500 hCPCs per well were seeded and incubated in growth media overnight.

    Protein Concentration:

    Article Title: TLR4 (toll-like receptor 4) activation suppresses autophagy through inhibition of FOXO3 and impairs phagocytic capacity of microglia
    Article Snippet: BCA protein assay reagents (Thermo Fisher Scientific, 23225) were used to measure protein concentration. .. After washing, proteins of interest were detected using either Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, 32106) or WesternBright ECL (Advansta, K-12045-D50).

    Article Title: Mitochondrial Uncoupling Protein 2 Knock-out Promotes Mitophagy to Decrease Retinal Ganglion Cell Death in a Mouse Model of Glaucoma
    Article Snippet: Protein concentration was estimated with a DC protein assay (Bio-Rad, catalog #5000111); 20 μg retinal homogenate was loaded on a 4%–12% Criterion TGX polyacrylamide gel (Bio-Rad, catalog #5678124) and run at 240 V for 35 min before semidry transfer to a 0.45 μm nitrocellulose membrane (Bio-Rad, catalog #1620115) at 16 V for 40 min. Leftover protein was visualized with Coomassie Blue staining, and the sum of Coomassie-labeled histone bands was used for protein normalization. .. Membranes were washed in TBS-T and incubated in secondary antibody for 2 h at room temperature before generation of chemiluminescent bands with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, catalog #32106) and visualization on photosensitive films (Thermo Fisher Scientific, catalog #34091).

    Article Title: SIV-Induced Immune Activation and Metabolic Alterations in the Dorsal Root Ganglia During Acute Infection
    Article Snippet: Protein concentration was estimated using the BioRad Protein Assay Kit (BioRad Laboratories, Hercules, CA) according to manufacturer instructions. .. Membranes were then washed in TBST and developed with Pierce ECL Western Blotting Substrate (Thermo Fisher) prior to exposure to radiography film.

    Article Title: Inactivation of mTORC2 in macrophages is a signature of colorectal cancer that promotes tumorigenesis
    Article Snippet: Protein concentration was adjusted with lysis buffer and 4× SDS reducing sample buffer containing 250 mM Tris-HCl (pH 6.8), 40% glycerol (Merck), 8% SDS (MilliporeSigma), 400 mM dithiothreitol (MilliporeSigma), and bromphenolblue (Merck) was added. .. Proteins were detected by chemiluminescence using the Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific).

    Sonication:

    Article Title: SIV-Induced Immune Activation and Metabolic Alterations in the Dorsal Root Ganglia During Acute Infection
    Article Snippet: Homogenates were then sonicated (3 times for 10 seconds) and centrifuged to remove insoluble material. .. Membranes were then washed in TBST and developed with Pierce ECL Western Blotting Substrate (Thermo Fisher) prior to exposure to radiography film.

    Affinity Purification:

    Article Title: Comparative flavivirus-host protein interaction mapping reveals mechanisms of dengue and Zika virus pathogenesis
    Article Snippet: C-terminally Strep II-tagged flavivirus NS5 constructs were expressed in HEK 293T and affinity purified as described for viral proteins in Expression and affinity purification of viral proteins . .. Signal was detected using Pierce ECL Western Blotting Substrate (Thermo Fisher).

    Binding Assay:

    Article Title: Anti‐inflammatory effects of miR‐21 in the macrophage response to peritonitis
    Article Snippet: Nonspecific binding was blocked with 7% nonfat milk in TBST for 1 h at room temperature. .. Pierce ECL Western blotting substrate (Thermo Fisher) was applied to the membrane as per the manufacturer's instructions.

    Magnetic Beads:

    Article Title: Tumor suppression of novel anti–PD-1 antibodies mediated through CD28 costimulatory pathway
    Article Snippet: Magnetic beads were attracted to the side of each tube with a magnet and washed three times with cell lysis buffer and then once with lysis buffer without detergent. .. Immunoblotting was performed by staining with the primary Ab overnight at 4°C followed by washing steps and then incubation with the HRP-conjugated secondary Ab and detection with the Pierce ECL Western blotting substrate (Thermo Fisher Scientific) in accordance with the manufacturer’s protocol.

    Isolation:

    Article Title: SIV-Induced Immune Activation and Metabolic Alterations in the Dorsal Root Ganglia During Acute Infection
    Article Snippet: Paragraph title: Protein Isolation and Western Blotting ... Membranes were then washed in TBST and developed with Pierce ECL Western Blotting Substrate (Thermo Fisher) prior to exposure to radiography film.

    Article Title: Anti‐inflammatory effects of miR‐21 in the macrophage response to peritonitis
    Article Snippet: Paragraph title: Protein isolation and Western blot ... Pierce ECL Western blotting substrate (Thermo Fisher) was applied to the membrane as per the manufacturer's instructions.

    Transfection:

    Article Title: Ripor2 is involved in auditory hair cell stereociliary bundle structure and orientation
    Article Snippet: Paragraph title: Transfection, Immunoprecipitation, Co-immunoprecipitation and Western Blot ... The results were visualized by chemiluminescence using Pierce ECL Western Blotting Substrate (Thermo Scientific).

    Avidin-Biotin Assay:

    Article Title: Tumor suppression of novel anti–PD-1 antibodies mediated through CD28 costimulatory pathway
    Article Snippet: For each condition, 10 million Jurkat cells stably expressing PD-1 (BPS Bioscience) were either left unstimulated or stimulated with biotinylated anti-CD3 (5 µg/ml; OKT3; eBioscience) and anti-CD28 (5 µg/ml; CD28.2; eBioscience) then aggregated with the addition of 50 µg/ml of avidin (Invitrogen). .. Immunoblotting was performed by staining with the primary Ab overnight at 4°C followed by washing steps and then incubation with the HRP-conjugated secondary Ab and detection with the Pierce ECL Western blotting substrate (Thermo Fisher Scientific) in accordance with the manufacturer’s protocol.

    Immunodetection:

    Article Title: Ripor2 is involved in auditory hair cell stereociliary bundle structure and orientation
    Article Snippet: Immunodetection was performed using anti-HA (Abcam) and anti-Myc (Cell Signaling) as primary mouse antibodies. .. The results were visualized by chemiluminescence using Pierce ECL Western Blotting Substrate (Thermo Scientific).

    Stripping:

    Article Title: SIV-Induced Immune Activation and Metabolic Alterations in the Dorsal Root Ganglia During Acute Infection
    Article Snippet: Membranes were then washed in TBST and developed with Pierce ECL Western Blotting Substrate (Thermo Fisher) prior to exposure to radiography film. .. Because GS, βIII tubulin, and β-actin have similar molecular weights, the membranes were first probed for GS, stripped using Restore PLUS Western Blot Stripping Buffer (Thermo Fisher) for 15 minutes at room temperature, then re-probed for βIII-tubulin, followed by β-actin.

    Construct:

    Article Title: Comparative flavivirus-host protein interaction mapping reveals mechanisms of dengue and Zika virus pathogenesis
    Article Snippet: C-terminally Strep II-tagged flavivirus NS5 constructs were expressed in HEK 293T and affinity purified as described for viral proteins in Expression and affinity purification of viral proteins . .. Signal was detected using Pierce ECL Western Blotting Substrate (Thermo Fisher).

    Staining:

    Article Title: Tumor suppression of novel anti–PD-1 antibodies mediated through CD28 costimulatory pathway
    Article Snippet: .. Immunoblotting was performed by staining with the primary Ab overnight at 4°C followed by washing steps and then incubation with the HRP-conjugated secondary Ab and detection with the Pierce ECL Western blotting substrate (Thermo Fisher Scientific) in accordance with the manufacturer’s protocol. .. Blots were imaged with the Fusion FX Vilber Lurmat camera (Witec), and the density of individual bands was evaluated using ImageJ software.

    Article Title: Mitochondrial Uncoupling Protein 2 Knock-out Promotes Mitophagy to Decrease Retinal Ganglion Cell Death in a Mouse Model of Glaucoma
    Article Snippet: Protein concentration was estimated with a DC protein assay (Bio-Rad, catalog #5000111); 20 μg retinal homogenate was loaded on a 4%–12% Criterion TGX polyacrylamide gel (Bio-Rad, catalog #5678124) and run at 240 V for 35 min before semidry transfer to a 0.45 μm nitrocellulose membrane (Bio-Rad, catalog #1620115) at 16 V for 40 min. Leftover protein was visualized with Coomassie Blue staining, and the sum of Coomassie-labeled histone bands was used for protein normalization. .. Membranes were washed in TBS-T and incubated in secondary antibody for 2 h at room temperature before generation of chemiluminescent bands with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, catalog #32106) and visualization on photosensitive films (Thermo Fisher Scientific, catalog #34091).

    Purification:

    Article Title: Comparative flavivirus-host protein interaction mapping reveals mechanisms of dengue and Zika virus pathogenesis
    Article Snippet: Paragraph title: NS5-PAF1C purification and Western blot ... Signal was detected using Pierce ECL Western Blotting Substrate (Thermo Fisher).

    SDS Page:

    Article Title: Akap1 genetic deletion increases the severity of hyperoxia-induced acute lung injury in mice
    Article Snippet: Equal amounts of protein (20 µg) were subjected SDS-PAGE by using 4–20% Tris-glycine gels (Bio-Rad), followed by electrotransfer to PVDF membrane. .. The proteins were visualized using Pierce ECL Western blotting substrate (Thermo Fisher Scientifics).

    Article Title: Effects and Mechanism of Nicotinamide Against UVA- and/or UVB-mediated DNA Damages in Normal Melanocytes
    Article Snippet: After UV exposure, cells were washed again and treated with 50 μm nicotinamide for 24 h. Cell lysates were prepared in ice-cold RIPA lysis buffer and subjected to SDS-PAGE. .. Proteins were detected by chemiluminescence using Pierce ECL Western Blotting Substrate, or SuperSignal West Femto Chemiluminescent Substrates (Pierce, Rockford, IL).

    Article Title: Accelerated decline in cognition in a mouse model of increased oxidative stress
    Article Snippet: For the detection of 4-HNE modified proteins, equal amounts of protein (40 μg/lane) were separated by SDS–PAGE, transferred to polyvinylidene difluoride membranes and treated with 250 mM sodium borohydride in 100 mM (3-(N-morpholino)propanesulfonic acid, MOPS), pH 8.0 for 15 min. .. This was followed by incubation with anti-rabbit IgG HRP conjugated antibody and the blot was developed using Pierce ECL Western Blotting Substrate (ThermoFisher Scientific, Waltham, MA).

    Article Title: Envelope Protein Glycosylation Mediates Zika Virus Pathogenesis
    Article Snippet: PNGaseF digestions were separated by SDS-PAGE (7.5% precast polyacrylamide gels; Bio-Rad) under nonreducing conditions and analyzed by Western or lectin blotting. .. Pierce ECL Western blotting substrate was used to detect HRP (Thermo).

    Article Title: Inactivation of mTORC2 in macrophages is a signature of colorectal cancer that promotes tumorigenesis
    Article Snippet: Samples were boiled for 5 minutes, resolved on 12% SDS-PAGE, and transferred to a nitrocellulose membrane (Peqlab). .. Proteins were detected by chemiluminescence using the Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific).

    Software:

    Article Title: Akap1 genetic deletion increases the severity of hyperoxia-induced acute lung injury in mice
    Article Snippet: The proteins were visualized using Pierce ECL Western blotting substrate (Thermo Fisher Scientifics). .. Protein bands were scanned, and densitometry was analyzed with NIH ImageJ software.

    Article Title: Tumor suppression of novel anti–PD-1 antibodies mediated through CD28 costimulatory pathway
    Article Snippet: Immunoblotting was performed by staining with the primary Ab overnight at 4°C followed by washing steps and then incubation with the HRP-conjugated secondary Ab and detection with the Pierce ECL Western blotting substrate (Thermo Fisher Scientific) in accordance with the manufacturer’s protocol. .. Blots were imaged with the Fusion FX Vilber Lurmat camera (Witec), and the density of individual bands was evaluated using ImageJ software.

    Article Title: Ponatinib shows potent antitumor activity in small cell carcinoma of the ovary hypercalcemic type (SCCOHT) through multi-kinase inhibition
    Article Snippet: .. Arrays were developed using Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific), and images were quantified using ImageJ software (NIH) using built-in gel quantification and background subtraction tools. ..

    Article Title: Accelerated decline in cognition in a mouse model of increased oxidative stress
    Article Snippet: This was followed by incubation with anti-rabbit IgG HRP conjugated antibody and the blot was developed using Pierce ECL Western Blotting Substrate (ThermoFisher Scientific, Waltham, MA). .. Images were taken using a G:BOX imaging system (Syngene, Frederick, MD) and quantified using ImageJ software (U.S. National Institutes of Health, Bethesda, MD, USA).

    Article Title: Decellularized Extracellular Matrix Hydrogels as a Delivery Platform for MicroRNA and Extracellular Vesicle Therapeutics
    Article Snippet: .. Prior to imaging, the membrane was incubated with Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific) for 1 minute and imaged with the Bio-Rad ChemiDoc MP System using Image Lab 3.0 software. .. A 96-well plate was coated with 0.1% porcine gelatin and 7500 hCPCs per well were seeded and incubated in growth media overnight.

    Article Title: SIV-Induced Immune Activation and Metabolic Alterations in the Dorsal Root Ganglia During Acute Infection
    Article Snippet: Membranes were then washed in TBST and developed with Pierce ECL Western Blotting Substrate (Thermo Fisher) prior to exposure to radiography film. .. Films were digitally scanned, and densitometry analysis was performed using ImageQuant TL 7.0 software (GE Life Sciences, Marlborough, MA).

    SDS-Gel:

    Article Title: TLR4 (toll-like receptor 4) activation suppresses autophagy through inhibition of FOXO3 and impairs phagocytic capacity of microglia
    Article Snippet: Samples were separated by SDS gel electrophoresis and electro-transferred onto polyvinylidene difluoride membrane in a semi-dry electrophoretic transfer cell (Bio-Rad). .. After washing, proteins of interest were detected using either Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, 32106) or WesternBright ECL (Advansta, K-12045-D50).

    Protein Extraction:

    Article Title: Akap1 genetic deletion increases the severity of hyperoxia-induced acute lung injury in mice
    Article Snippet: Frozen lung tissue was pulverized and lysed in protein extraction buffer (20 mM Tris·HCl, pH 7.4, 150 mM NaCl, and 0.5% Triton X-100), and the supernatant, containing soluble protein, was collected by high-speed centrifugation (14,000 g ) for 15 min at 4°C. .. The proteins were visualized using Pierce ECL Western blotting substrate (Thermo Fisher Scientifics).

    Immunoprecipitation:

    Article Title: Tumor suppression of novel anti–PD-1 antibodies mediated through CD28 costimulatory pathway
    Article Snippet: Paragraph title: Immunoprecipitation of the cellular PD-1 protein complex ... Immunoblotting was performed by staining with the primary Ab overnight at 4°C followed by washing steps and then incubation with the HRP-conjugated secondary Ab and detection with the Pierce ECL Western blotting substrate (Thermo Fisher Scientific) in accordance with the manufacturer’s protocol.

    Article Title: Ripor2 is involved in auditory hair cell stereociliary bundle structure and orientation
    Article Snippet: Paragraph title: Transfection, Immunoprecipitation, Co-immunoprecipitation and Western Blot ... The results were visualized by chemiluminescence using Pierce ECL Western Blotting Substrate (Thermo Scientific).

    Marker:

    Article Title: Ripor2 is involved in auditory hair cell stereociliary bundle structure and orientation
    Article Snippet: Samples were boiled with a lane marker containing reducing agent at 100°C for 5 min. For western blots pulldown samples were loaded onto precast 4–20% gradient SDS-polyacrylamide gels (Lonza) and transferred to a 0.2-μm PVDF membrane (Bio-Rad). .. The results were visualized by chemiluminescence using Pierce ECL Western Blotting Substrate (Thermo Scientific).

    Lysis:

    Article Title: TLR4 (toll-like receptor 4) activation suppresses autophagy through inhibition of FOXO3 and impairs phagocytic capacity of microglia
    Article Snippet: Cells were harvested and lysed in lysis buffer containing 250 mM sucrose (Duchefa Biochemie, 57-50-1), 50 mM NaCl, 20 mM Tris-HCl, pH 7.4, 1 mM EDTA, 1% Triton X-100, 1× protease and phosphate inhibitor cocktail (Thermo Fisher Scientific, 78440), 1 mM dithiothreitol, and 1 mM phenylmethylsulfonyl fluoride for 15 min on ice. .. After washing, proteins of interest were detected using either Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific, 32106) or WesternBright ECL (Advansta, K-12045-D50).

    Article Title: Tumor suppression of novel anti–PD-1 antibodies mediated through CD28 costimulatory pathway
    Article Snippet: Magnetic beads were attracted to the side of each tube with a magnet and washed three times with cell lysis buffer and then once with lysis buffer without detergent. .. Immunoblotting was performed by staining with the primary Ab overnight at 4°C followed by washing steps and then incubation with the HRP-conjugated secondary Ab and detection with the Pierce ECL Western blotting substrate (Thermo Fisher Scientific) in accordance with the manufacturer’s protocol.

    Article Title: Ripor2 is involved in auditory hair cell stereociliary bundle structure and orientation
    Article Snippet: Beads were washed twice with the manufacturer’s immunoprecipitation lysis and wash buffer. .. The results were visualized by chemiluminescence using Pierce ECL Western Blotting Substrate (Thermo Scientific).

    Article Title: Effects and Mechanism of Nicotinamide Against UVA- and/or UVB-mediated DNA Damages in Normal Melanocytes
    Article Snippet: After UV exposure, cells were washed again and treated with 50 μm nicotinamide for 24 h. Cell lysates were prepared in ice-cold RIPA lysis buffer and subjected to SDS-PAGE. .. Proteins were detected by chemiluminescence using Pierce ECL Western Blotting Substrate, or SuperSignal West Femto Chemiluminescent Substrates (Pierce, Rockford, IL).

    Article Title: Accelerated decline in cognition in a mouse model of increased oxidative stress
    Article Snippet: For Western blotting, one hippocampus was homogenized in RIPA lysis and extraction buffer (ThermoFisher Scientific, Waltham, MA). .. This was followed by incubation with anti-rabbit IgG HRP conjugated antibody and the blot was developed using Pierce ECL Western Blotting Substrate (ThermoFisher Scientific, Waltham, MA).

    Article Title: Inactivation of mTORC2 in macrophages is a signature of colorectal cancer that promotes tumorigenesis
    Article Snippet: Protein concentration was adjusted with lysis buffer and 4× SDS reducing sample buffer containing 250 mM Tris-HCl (pH 6.8), 40% glycerol (Merck), 8% SDS (MilliporeSigma), 400 mM dithiothreitol (MilliporeSigma), and bromphenolblue (Merck) was added. .. Proteins were detected by chemiluminescence using the Pierce ECL Western Blotting Substrate (Thermo Fisher Scientific).

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  • 90
    Thermo Fisher pierce ecl western blotting substrate
    Pierce Ecl Western Blotting Substrate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1238 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pierce ecl western blotting substrate/product/Thermo Fisher
    Average 90 stars, based on 1238 article reviews
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    pierce ecl western blotting substrate - by Bioz Stars, 2020-01
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