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Cell Signaling Technology Inc protease inhibitor cocktail
Protease Inhibitor Cocktail, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 87 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 99 stars, based on 87 article reviews
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Centrifugation:

Article Title: BRD4-Regulated Molecular Targets in Mantle Cell Lymphoma: Insights into Targeted Therapeutic Approach
Article Snippet: Cross-linked cells were lysed with buffer A supplemented with a dithiothreitol and protease inhibitor cocktail, and the nuclear fraction was collected and treated with micrococcal nuclease in buffer B containing dithiothreitol at 37˚C for 20 min. After sonication in 1x ChIP buffer supplemented with a protease inhibitor cocktail, lysates were clarified by centrifugation at 9,300 g for 10 min at 4˚C. .. Chromatin samples were diluted into five-fold in ChIP buffer containing a protease inhibitor cocktail, and incubated with either anti-BRD4 antibody (E2A7X, Cell Signaling Technology) or anti-IgG antibody (background control) overnight at 4˚C.

Article Title: Novel replisome-associated proteins at cellular replication forks in EBV-transformed B lymphocytes
Article Snippet: After centrifugation, nuclei were resuspended in cold lysis buffer (1% SDS, 50 mM Tris, pH 8.0), sonicated using a microtip sonicator at 8W with 10 sec on and 20 sec off pulses on ice three times to break nuclear membranes. .. Samples were centrifuged at 16,100 × g for 10 min at 4°C to remove debris and diluted with the same volume of ice-cold PBS containing protease inhibitor cocktail (Cell Signaling, #5871), and incubated with pre-washed 150 μl of streptavidin agarose beads (EMD Millipore, #69203) overnight at 4°C.

Blocking Assay:

Article Title: Obesity, Diabetes and Energy Homeostasis: Reduced uterine perfusion pressure causes loss of pancreatic β-cell area but normal function in fetal rat offspring
Article Snippet: Placenta or pancreatic lysates were prepared by freezing with liquid nitrogen, processing with mortar and pestle, and then resuspending in 1× radioimmunoprecipitation assay lysis (cat. no. 5871, Cell Signaling) buffer with protease inhibitor cocktail (Cell Signaling). .. After transfer to a polyvinylidene fluoride membrane, it was blocked with Tris-buffered saline blocking solution (LI-COR Biosciences) or 5% nonfat dried milk (RPI) in PBS for 30 min and incubated overnight at 4°C with primary antibodies: mammalian target of rapamycin (mTOR) (Cell Signaling), O -GlcNAc transferase (OGT) (Cell Signaling), phospho-S6 (Ser240 ; Cell Signaling), S6 (Cell Signaling), phospho-Akt (Ser473 ; Cell Signaling), RL2 (Abcam), β-actin (Cell Signaling), and Vinculin (Cell Signaling).

SYBR Green Assay:

Article Title: The Dietary Flavone Luteolin Epigenetically Activates the Nrf2 Pathway and Blocks Cell Transformation in Human Colorectal Cancer HCT116 Cells
Article Snippet: Power SYBR Green PCR Master Mix was purchased from Applied Biosystems (Carlsbad, CA, USA). .. Protease inhibitor cocktail, radioimmunoprecipitation assay (RIPA) buffer, and antibodies against HO-1, NQO1, β-actin, HDACs (HDAC1-7) and DNMTs (DNMT1, DNMT3a/b) were supplied by Cell Signaling Technology (Beverly, MA, USA).

Incubation:

Article Title: BRD4-Regulated Molecular Targets in Mantle Cell Lymphoma: Insights into Targeted Therapeutic Approach
Article Snippet: .. Chromatin samples were diluted into five-fold in ChIP buffer containing a protease inhibitor cocktail, and incubated with either anti-BRD4 antibody (E2A7X, Cell Signaling Technology) or anti-IgG antibody (background control) overnight at 4˚C. .. The antibody-bound complexes were captured by incubation with protein G magnetic beads for 2 h at 4˚C and washed in low and high salt ChIP buffer.

Article Title: Obesity, Diabetes and Energy Homeostasis: Reduced uterine perfusion pressure causes loss of pancreatic β-cell area but normal function in fetal rat offspring
Article Snippet: Placenta or pancreatic lysates were prepared by freezing with liquid nitrogen, processing with mortar and pestle, and then resuspending in 1× radioimmunoprecipitation assay lysis (cat. no. 5871, Cell Signaling) buffer with protease inhibitor cocktail (Cell Signaling). .. After transfer to a polyvinylidene fluoride membrane, it was blocked with Tris-buffered saline blocking solution (LI-COR Biosciences) or 5% nonfat dried milk (RPI) in PBS for 30 min and incubated overnight at 4°C with primary antibodies: mammalian target of rapamycin (mTOR) (Cell Signaling), O -GlcNAc transferase (OGT) (Cell Signaling), phospho-S6 (Ser240 ; Cell Signaling), S6 (Cell Signaling), phospho-Akt (Ser473 ; Cell Signaling), RL2 (Abcam), β-actin (Cell Signaling), and Vinculin (Cell Signaling).

Article Title: Novel replisome-associated proteins at cellular replication forks in EBV-transformed B lymphocytes
Article Snippet: .. Samples were centrifuged at 16,100 × g for 10 min at 4°C to remove debris and diluted with the same volume of ice-cold PBS containing protease inhibitor cocktail (Cell Signaling, #5871), and incubated with pre-washed 150 μl of streptavidin agarose beads (EMD Millipore, #69203) overnight at 4°C. .. Protein-DNA complexes were either analyzed by mass spectrometry or eluted with 70 μl of 2 × Laemli buffer and boiled at 95°C for 25 min, followed by immunoblotting.

Article Title: High Fat Diet Affects Pre-Gestational Adiposity and Glucose Tolerance Perturbing Gestational Placental Macronutrient Transporters Culminating in an Obese Offspring in Wild type and Glucose Transporter Isoform 3 Heterozygous Null Mice
Article Snippet: Once cells were confluent, they were washed twice with PBS then incubated in low glucose (2.5 mM) over varying durations. .. These cells were then lysed in cell lysis buffer (Cell Signaling Technology, Denvers, MA), supplemented with 1 % phosphatase and protease inhibitor cocktail (Cell Signaling Technology, Danvers, MA).

Mass Spectrometry:

Article Title: Novel replisome-associated proteins at cellular replication forks in EBV-transformed B lymphocytes
Article Snippet: Samples were centrifuged at 16,100 × g for 10 min at 4°C to remove debris and diluted with the same volume of ice-cold PBS containing protease inhibitor cocktail (Cell Signaling, #5871), and incubated with pre-washed 150 μl of streptavidin agarose beads (EMD Millipore, #69203) overnight at 4°C. .. Protein-DNA complexes were either analyzed by mass spectrometry or eluted with 70 μl of 2 × Laemli buffer and boiled at 95°C for 25 min, followed by immunoblotting.

BIA-KA:

Article Title: Endoplasmic Reticulum Protein Disulfide Isomerase Shapes T Cell Efficacy for Adoptive Cellular Therapy of Tumors
Article Snippet: Immunoblotting T cells were lysed in RIPA buffer (Sigma, St. Louis, MO, USA) supplemented with protease inhibitor cocktail (Cell Signaling Technology, Danvers, MA, USA) and phosphatase inhibitors I and II (Sigma, St. Louis, MO, USA). .. Protein concentrations were normalized using Pierce BCA Kit (Thermo Fisher, Waltham, MA, USA) and loaded to 4%–10% agarose gels (BioRad, Hercules, CA, USA).

Article Title: Obesity, Diabetes and Energy Homeostasis: Reduced uterine perfusion pressure causes loss of pancreatic β-cell area but normal function in fetal rat offspring
Article Snippet: Placenta or pancreatic lysates were prepared by freezing with liquid nitrogen, processing with mortar and pestle, and then resuspending in 1× radioimmunoprecipitation assay lysis (cat. no. 5871, Cell Signaling) buffer with protease inhibitor cocktail (Cell Signaling). .. Samples were sonicated (Fisher Scientific Sonic Dismembrator model 500; 10 × 3 s, 30% amplitude) and centrifuged to remove the supernatant for total protein quantification via the Pierce BCA Protein Assay Kit (Thermo Scientific), according to the manufacturer’s instructions.

Western Blot:

Article Title: RANKL-mediated osteoclastogenic differentiation of macrophages in the abdominal aorta of angiotensin II-infused apolipoprotein E knockout mice
Article Snippet: Paragraph title: Western blotting. ... Protein extraction from cultured cells and isolated aortas was performed at 0°C to 4°C with a radioimmunoprecipitation assay buffer and protease inhibitor cocktail (Cell Signaling Technology).

Article Title: Astaxanthin Suppresses Cigarette Smoke-Induced Emphysema through Nrf2 Activation in Mice
Article Snippet: Paragraph title: 4.5. Western Blot Analysis ... Approximately 30 mg of the lung sample was soaked in 300 µL of radioimmunoprecipitation assay (RIPA) buffer (Beyotime Biotechnology, Shanghai, China) supplemented with the protease inhibitor phenylmethanesulfonyl fluoride (Beyotime Biotechnology, Shanghai, China) and Protease Inhibitor Cocktail (Cell Signaling Technology Japan, Tokyo, Japan).

Article Title: Obesity, Diabetes and Energy Homeostasis: Reduced uterine perfusion pressure causes loss of pancreatic β-cell area but normal function in fetal rat offspring
Article Snippet: Paragraph title: Protein isolation and Western blot. ... Placenta or pancreatic lysates were prepared by freezing with liquid nitrogen, processing with mortar and pestle, and then resuspending in 1× radioimmunoprecipitation assay lysis (cat. no. 5871, Cell Signaling) buffer with protease inhibitor cocktail (Cell Signaling).

Article Title: High Fat Diet Affects Pre-Gestational Adiposity and Glucose Tolerance Perturbing Gestational Placental Macronutrient Transporters Culminating in an Obese Offspring in Wild type and Glucose Transporter Isoform 3 Heterozygous Null Mice
Article Snippet: These cells were then lysed in cell lysis buffer (Cell Signaling Technology, Denvers, MA), supplemented with 1 % phosphatase and protease inhibitor cocktail (Cell Signaling Technology, Danvers, MA). .. The cell lysates (30 μg) were subjected to Western Blot analysis as described above.

Transfection:

Article Title: Environmental control of astrocyte pathogenic activities in CNS inflammation
Article Snippet: HEK293 cells were transfected with human WT IRE1α (a gift from Fumihiko Urano, Addgene plasmid #20744) , Sigmar1-myc-FLAG transcript variant 1 (#RC201206, Origene), or Sigmar1-myc-FLAG transcript variant 2 (#RC214331, Origene) using Lipofectamine 2000 according to the manufacturer’s protocol. .. 16 hours later, cells were lysed in ice cold non-denaturing lysis buffer (20 mM Tris-HCl pH=8.0, 137 mM NaCl, 1% NP-40, 2 mM EDTA, recipe obtained from Abcam) supplemented with protease inhibitor cocktail (#5871, Cell Signaling, 1:100 working concentration) and phosphatase inhibitor cocktail (#5870, Cell Signaling, 1:100 working concentration).

Immunoprecipitation:

Article Title: BRD4-Regulated Molecular Targets in Mantle Cell Lymphoma: Insights into Targeted Therapeutic Approach
Article Snippet: Chromatin samples were diluted into five-fold in ChIP buffer containing a protease inhibitor cocktail, and incubated with either anti-BRD4 antibody (E2A7X, Cell Signaling Technology) or anti-IgG antibody (background control) overnight at 4˚C. .. The immunoprecipitated DNA was purified using a spin column.

Article Title: Environmental control of astrocyte pathogenic activities in CNS inflammation
Article Snippet: 16 hours later, cells were lysed in ice cold non-denaturing lysis buffer (20 mM Tris-HCl pH=8.0, 137 mM NaCl, 1% NP-40, 2 mM EDTA, recipe obtained from Abcam) supplemented with protease inhibitor cocktail (#5871, Cell Signaling, 1:100 working concentration) and phosphatase inhibitor cocktail (#5870, Cell Signaling, 1:100 working concentration). .. Immunoprecipitation for c-myc was performed for 4 hours at 4C using the following kit (#23620, Thermo Fisher Scientific).

Protease Inhibitor:

Article Title: Endoplasmic Reticulum Protein Disulfide Isomerase Shapes T Cell Efficacy for Adoptive Cellular Therapy of Tumors
Article Snippet: .. Immunoblotting T cells were lysed in RIPA buffer (Sigma, St. Louis, MO, USA) supplemented with protease inhibitor cocktail (Cell Signaling Technology, Danvers, MA, USA) and phosphatase inhibitors I and II (Sigma, St. Louis, MO, USA). .. Protein concentrations were normalized using Pierce BCA Kit (Thermo Fisher, Waltham, MA, USA) and loaded to 4%–10% agarose gels (BioRad, Hercules, CA, USA).

Article Title: RANKL-mediated osteoclastogenic differentiation of macrophages in the abdominal aorta of angiotensin II-infused apolipoprotein E knockout mice
Article Snippet: .. Protein extraction from cultured cells and isolated aortas was performed at 0°C to 4°C with a radioimmunoprecipitation assay buffer and protease inhibitor cocktail (Cell Signaling Technology). .. Twenty-microgram samples were separated by electrophoresis on 8% to 12% polyacrylamide gels in Laemmli buffer and transferred onto polyvinylidene difluoride membranes.

Article Title: Astaxanthin Suppresses Cigarette Smoke-Induced Emphysema through Nrf2 Activation in Mice
Article Snippet: .. Approximately 30 mg of the lung sample was soaked in 300 µL of radioimmunoprecipitation assay (RIPA) buffer (Beyotime Biotechnology, Shanghai, China) supplemented with the protease inhibitor phenylmethanesulfonyl fluoride (Beyotime Biotechnology, Shanghai, China) and Protease Inhibitor Cocktail (Cell Signaling Technology Japan, Tokyo, Japan). ..

Article Title: The Dietary Flavone Luteolin Epigenetically Activates the Nrf2 Pathway and Blocks Cell Transformation in Human Colorectal Cancer HCT116 Cells
Article Snippet: .. Protease inhibitor cocktail, radioimmunoprecipitation assay (RIPA) buffer, and antibodies against HO-1, NQO1, β-actin, HDACs (HDAC1-7) and DNMTs (DNMT1, DNMT3a/b) were supplied by Cell Signaling Technology (Beverly, MA, USA). ..

Article Title: BRD4-Regulated Molecular Targets in Mantle Cell Lymphoma: Insights into Targeted Therapeutic Approach
Article Snippet: .. Chromatin samples were diluted into five-fold in ChIP buffer containing a protease inhibitor cocktail, and incubated with either anti-BRD4 antibody (E2A7X, Cell Signaling Technology) or anti-IgG antibody (background control) overnight at 4˚C. .. The antibody-bound complexes were captured by incubation with protein G magnetic beads for 2 h at 4˚C and washed in low and high salt ChIP buffer.

Article Title: Obesity, Diabetes and Energy Homeostasis: Reduced uterine perfusion pressure causes loss of pancreatic β-cell area but normal function in fetal rat offspring
Article Snippet: .. Placenta or pancreatic lysates were prepared by freezing with liquid nitrogen, processing with mortar and pestle, and then resuspending in 1× radioimmunoprecipitation assay lysis (cat. no. 5871, Cell Signaling) buffer with protease inhibitor cocktail (Cell Signaling). .. Samples were sonicated (Fisher Scientific Sonic Dismembrator model 500; 10 × 3 s, 30% amplitude) and centrifuged to remove the supernatant for total protein quantification via the Pierce BCA Protein Assay Kit (Thermo Scientific), according to the manufacturer’s instructions.

Article Title: Environmental control of astrocyte pathogenic activities in CNS inflammation
Article Snippet: .. 16 hours later, cells were lysed in ice cold non-denaturing lysis buffer (20 mM Tris-HCl pH=8.0, 137 mM NaCl, 1% NP-40, 2 mM EDTA, recipe obtained from Abcam) supplemented with protease inhibitor cocktail (#5871, Cell Signaling, 1:100 working concentration) and phosphatase inhibitor cocktail (#5870, Cell Signaling, 1:100 working concentration). .. Immunoprecipitation for c-myc was performed for 4 hours at 4C using the following kit (#23620, Thermo Fisher Scientific).

Article Title: Novel replisome-associated proteins at cellular replication forks in EBV-transformed B lymphocytes
Article Snippet: .. Samples were centrifuged at 16,100 × g for 10 min at 4°C to remove debris and diluted with the same volume of ice-cold PBS containing protease inhibitor cocktail (Cell Signaling, #5871), and incubated with pre-washed 150 μl of streptavidin agarose beads (EMD Millipore, #69203) overnight at 4°C. .. Protein-DNA complexes were either analyzed by mass spectrometry or eluted with 70 μl of 2 × Laemli buffer and boiled at 95°C for 25 min, followed by immunoblotting.

Article Title: High Fat Diet Affects Pre-Gestational Adiposity and Glucose Tolerance Perturbing Gestational Placental Macronutrient Transporters Culminating in an Obese Offspring in Wild type and Glucose Transporter Isoform 3 Heterozygous Null Mice
Article Snippet: .. These cells were then lysed in cell lysis buffer (Cell Signaling Technology, Denvers, MA), supplemented with 1 % phosphatase and protease inhibitor cocktail (Cell Signaling Technology, Danvers, MA). .. The cell lysates (30 μg) were subjected to Western Blot analysis as described above.

Cell Culture:

Article Title: RANKL-mediated osteoclastogenic differentiation of macrophages in the abdominal aorta of angiotensin II-infused apolipoprotein E knockout mice
Article Snippet: .. Protein extraction from cultured cells and isolated aortas was performed at 0°C to 4°C with a radioimmunoprecipitation assay buffer and protease inhibitor cocktail (Cell Signaling Technology). .. Twenty-microgram samples were separated by electrophoresis on 8% to 12% polyacrylamide gels in Laemmli buffer and transferred onto polyvinylidene difluoride membranes.

Article Title: High Fat Diet Affects Pre-Gestational Adiposity and Glucose Tolerance Perturbing Gestational Placental Macronutrient Transporters Culminating in an Obese Offspring in Wild type and Glucose Transporter Isoform 3 Heterozygous Null Mice
Article Snippet: glut3 +/+ and glut3 +/− embryonic stem cells were cultured in Knock Out DMEM containing high glucose (25 mM) supplemented with 15% FBS and 1% penicillin-streptomycin in a humidified incubator at 37°C in an atmosphere of 5% CO2 and 95% air. .. These cells were then lysed in cell lysis buffer (Cell Signaling Technology, Denvers, MA), supplemented with 1 % phosphatase and protease inhibitor cocktail (Cell Signaling Technology, Danvers, MA).

Protein Concentration:

Article Title: Astaxanthin Suppresses Cigarette Smoke-Induced Emphysema through Nrf2 Activation in Mice
Article Snippet: Approximately 30 mg of the lung sample was soaked in 300 µL of radioimmunoprecipitation assay (RIPA) buffer (Beyotime Biotechnology, Shanghai, China) supplemented with the protease inhibitor phenylmethanesulfonyl fluoride (Beyotime Biotechnology, Shanghai, China) and Protease Inhibitor Cocktail (Cell Signaling Technology Japan, Tokyo, Japan). .. The supernatant was collected, and the protein concentration was determined with the colorimetric bicinchoninic acid protein assay kit (Pierce, Waltham, MA, USA) according to the manufacturer’s instructions.

Polymerase Chain Reaction:

Article Title: The Dietary Flavone Luteolin Epigenetically Activates the Nrf2 Pathway and Blocks Cell Transformation in Human Colorectal Cancer HCT116 Cells
Article Snippet: Power SYBR Green PCR Master Mix was purchased from Applied Biosystems (Carlsbad, CA, USA). .. Protease inhibitor cocktail, radioimmunoprecipitation assay (RIPA) buffer, and antibodies against HO-1, NQO1, β-actin, HDACs (HDAC1-7) and DNMTs (DNMT1, DNMT3a/b) were supplied by Cell Signaling Technology (Beverly, MA, USA).

Sonication:

Article Title: BRD4-Regulated Molecular Targets in Mantle Cell Lymphoma: Insights into Targeted Therapeutic Approach
Article Snippet: Cross-linked cells were lysed with buffer A supplemented with a dithiothreitol and protease inhibitor cocktail, and the nuclear fraction was collected and treated with micrococcal nuclease in buffer B containing dithiothreitol at 37˚C for 20 min. After sonication in 1x ChIP buffer supplemented with a protease inhibitor cocktail, lysates were clarified by centrifugation at 9,300 g for 10 min at 4˚C. .. Chromatin samples were diluted into five-fold in ChIP buffer containing a protease inhibitor cocktail, and incubated with either anti-BRD4 antibody (E2A7X, Cell Signaling Technology) or anti-IgG antibody (background control) overnight at 4˚C.

Article Title: Novel replisome-associated proteins at cellular replication forks in EBV-transformed B lymphocytes
Article Snippet: After centrifugation, nuclei were resuspended in cold lysis buffer (1% SDS, 50 mM Tris, pH 8.0), sonicated using a microtip sonicator at 8W with 10 sec on and 20 sec off pulses on ice three times to break nuclear membranes. .. Samples were centrifuged at 16,100 × g for 10 min at 4°C to remove debris and diluted with the same volume of ice-cold PBS containing protease inhibitor cocktail (Cell Signaling, #5871), and incubated with pre-washed 150 μl of streptavidin agarose beads (EMD Millipore, #69203) overnight at 4°C.

ChIP-sequencing:

Article Title: BRD4-Regulated Molecular Targets in Mantle Cell Lymphoma: Insights into Targeted Therapeutic Approach
Article Snippet: Chromatin immunoprecipitation with parallel sequencing (ChIP-Seq). .. Chromatin samples were diluted into five-fold in ChIP buffer containing a protease inhibitor cocktail, and incubated with either anti-BRD4 antibody (E2A7X, Cell Signaling Technology) or anti-IgG antibody (background control) overnight at 4˚C.

Magnetic Beads:

Article Title: BRD4-Regulated Molecular Targets in Mantle Cell Lymphoma: Insights into Targeted Therapeutic Approach
Article Snippet: ChIP was performed using the SimpleChIP Plus Enzymatic Chromatin IP Kit (Magnetic Beads; Cell Signaling Technology) according to the manufacturer’s protocol. .. Chromatin samples were diluted into five-fold in ChIP buffer containing a protease inhibitor cocktail, and incubated with either anti-BRD4 antibody (E2A7X, Cell Signaling Technology) or anti-IgG antibody (background control) overnight at 4˚C.

Isolation:

Article Title: RANKL-mediated osteoclastogenic differentiation of macrophages in the abdominal aorta of angiotensin II-infused apolipoprotein E knockout mice
Article Snippet: .. Protein extraction from cultured cells and isolated aortas was performed at 0°C to 4°C with a radioimmunoprecipitation assay buffer and protease inhibitor cocktail (Cell Signaling Technology). .. Twenty-microgram samples were separated by electrophoresis on 8% to 12% polyacrylamide gels in Laemmli buffer and transferred onto polyvinylidene difluoride membranes.

Article Title: Obesity, Diabetes and Energy Homeostasis: Reduced uterine perfusion pressure causes loss of pancreatic β-cell area but normal function in fetal rat offspring
Article Snippet: Paragraph title: Protein isolation and Western blot. ... Placenta or pancreatic lysates were prepared by freezing with liquid nitrogen, processing with mortar and pestle, and then resuspending in 1× radioimmunoprecipitation assay lysis (cat. no. 5871, Cell Signaling) buffer with protease inhibitor cocktail (Cell Signaling).

Article Title: Novel replisome-associated proteins at cellular replication forks in EBV-transformed B lymphocytes
Article Snippet: Paragraph title: Isolation of proteins on nascent DNA (iPOND) ... Samples were centrifuged at 16,100 × g for 10 min at 4°C to remove debris and diluted with the same volume of ice-cold PBS containing protease inhibitor cocktail (Cell Signaling, #5871), and incubated with pre-washed 150 μl of streptavidin agarose beads (EMD Millipore, #69203) overnight at 4°C.

Bicinchoninic Acid Protein Assay:

Article Title: Astaxanthin Suppresses Cigarette Smoke-Induced Emphysema through Nrf2 Activation in Mice
Article Snippet: Approximately 30 mg of the lung sample was soaked in 300 µL of radioimmunoprecipitation assay (RIPA) buffer (Beyotime Biotechnology, Shanghai, China) supplemented with the protease inhibitor phenylmethanesulfonyl fluoride (Beyotime Biotechnology, Shanghai, China) and Protease Inhibitor Cocktail (Cell Signaling Technology Japan, Tokyo, Japan). .. The supernatant was collected, and the protein concentration was determined with the colorimetric bicinchoninic acid protein assay kit (Pierce, Waltham, MA, USA) according to the manufacturer’s instructions.

Size-exclusion Chromatography:

Article Title: Novel replisome-associated proteins at cellular replication forks in EBV-transformed B lymphocytes
Article Snippet: After centrifugation, nuclei were resuspended in cold lysis buffer (1% SDS, 50 mM Tris, pH 8.0), sonicated using a microtip sonicator at 8W with 10 sec on and 20 sec off pulses on ice three times to break nuclear membranes. .. Samples were centrifuged at 16,100 × g for 10 min at 4°C to remove debris and diluted with the same volume of ice-cold PBS containing protease inhibitor cocktail (Cell Signaling, #5871), and incubated with pre-washed 150 μl of streptavidin agarose beads (EMD Millipore, #69203) overnight at 4°C.

Purification:

Article Title: BRD4-Regulated Molecular Targets in Mantle Cell Lymphoma: Insights into Targeted Therapeutic Approach
Article Snippet: Chromatin samples were diluted into five-fold in ChIP buffer containing a protease inhibitor cocktail, and incubated with either anti-BRD4 antibody (E2A7X, Cell Signaling Technology) or anti-IgG antibody (background control) overnight at 4˚C. .. The immunoprecipitated DNA was purified using a spin column.

Sequencing:

Article Title: BRD4-Regulated Molecular Targets in Mantle Cell Lymphoma: Insights into Targeted Therapeutic Approach
Article Snippet: Chromatin immunoprecipitation with parallel sequencing (ChIP-Seq). .. Chromatin samples were diluted into five-fold in ChIP buffer containing a protease inhibitor cocktail, and incubated with either anti-BRD4 antibody (E2A7X, Cell Signaling Technology) or anti-IgG antibody (background control) overnight at 4˚C.

Protein Extraction:

Article Title: RANKL-mediated osteoclastogenic differentiation of macrophages in the abdominal aorta of angiotensin II-infused apolipoprotein E knockout mice
Article Snippet: .. Protein extraction from cultured cells and isolated aortas was performed at 0°C to 4°C with a radioimmunoprecipitation assay buffer and protease inhibitor cocktail (Cell Signaling Technology). .. Twenty-microgram samples were separated by electrophoresis on 8% to 12% polyacrylamide gels in Laemmli buffer and transferred onto polyvinylidene difluoride membranes.

Polyacrylamide Gel Electrophoresis:

Article Title: Astaxanthin Suppresses Cigarette Smoke-Induced Emphysema through Nrf2 Activation in Mice
Article Snippet: Approximately 30 mg of the lung sample was soaked in 300 µL of radioimmunoprecipitation assay (RIPA) buffer (Beyotime Biotechnology, Shanghai, China) supplemented with the protease inhibitor phenylmethanesulfonyl fluoride (Beyotime Biotechnology, Shanghai, China) and Protease Inhibitor Cocktail (Cell Signaling Technology Japan, Tokyo, Japan). .. The supernatant was subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis with Mini-PROTEAN TGX Precast Protein Gels (4561023, Bio-Rad, Hercules, California, USA).

Chromatin Immunoprecipitation:

Article Title: BRD4-Regulated Molecular Targets in Mantle Cell Lymphoma: Insights into Targeted Therapeutic Approach
Article Snippet: .. Chromatin samples were diluted into five-fold in ChIP buffer containing a protease inhibitor cocktail, and incubated with either anti-BRD4 antibody (E2A7X, Cell Signaling Technology) or anti-IgG antibody (background control) overnight at 4˚C. .. The antibody-bound complexes were captured by incubation with protein G magnetic beads for 2 h at 4˚C and washed in low and high salt ChIP buffer.

Plasmid Preparation:

Article Title: Environmental control of astrocyte pathogenic activities in CNS inflammation
Article Snippet: HEK293 cells were transfected with human WT IRE1α (a gift from Fumihiko Urano, Addgene plasmid #20744) , Sigmar1-myc-FLAG transcript variant 1 (#RC201206, Origene), or Sigmar1-myc-FLAG transcript variant 2 (#RC214331, Origene) using Lipofectamine 2000 according to the manufacturer’s protocol. .. 16 hours later, cells were lysed in ice cold non-denaturing lysis buffer (20 mM Tris-HCl pH=8.0, 137 mM NaCl, 1% NP-40, 2 mM EDTA, recipe obtained from Abcam) supplemented with protease inhibitor cocktail (#5871, Cell Signaling, 1:100 working concentration) and phosphatase inhibitor cocktail (#5870, Cell Signaling, 1:100 working concentration).

Electrophoresis:

Article Title: RANKL-mediated osteoclastogenic differentiation of macrophages in the abdominal aorta of angiotensin II-infused apolipoprotein E knockout mice
Article Snippet: Protein extraction from cultured cells and isolated aortas was performed at 0°C to 4°C with a radioimmunoprecipitation assay buffer and protease inhibitor cocktail (Cell Signaling Technology). .. Twenty-microgram samples were separated by electrophoresis on 8% to 12% polyacrylamide gels in Laemmli buffer and transferred onto polyvinylidene difluoride membranes.

Radio Immunoprecipitation:

Article Title: RANKL-mediated osteoclastogenic differentiation of macrophages in the abdominal aorta of angiotensin II-infused apolipoprotein E knockout mice
Article Snippet: .. Protein extraction from cultured cells and isolated aortas was performed at 0°C to 4°C with a radioimmunoprecipitation assay buffer and protease inhibitor cocktail (Cell Signaling Technology). .. Twenty-microgram samples were separated by electrophoresis on 8% to 12% polyacrylamide gels in Laemmli buffer and transferred onto polyvinylidene difluoride membranes.

Article Title: Astaxanthin Suppresses Cigarette Smoke-Induced Emphysema through Nrf2 Activation in Mice
Article Snippet: .. Approximately 30 mg of the lung sample was soaked in 300 µL of radioimmunoprecipitation assay (RIPA) buffer (Beyotime Biotechnology, Shanghai, China) supplemented with the protease inhibitor phenylmethanesulfonyl fluoride (Beyotime Biotechnology, Shanghai, China) and Protease Inhibitor Cocktail (Cell Signaling Technology Japan, Tokyo, Japan). ..

Article Title: The Dietary Flavone Luteolin Epigenetically Activates the Nrf2 Pathway and Blocks Cell Transformation in Human Colorectal Cancer HCT116 Cells
Article Snippet: .. Protease inhibitor cocktail, radioimmunoprecipitation assay (RIPA) buffer, and antibodies against HO-1, NQO1, β-actin, HDACs (HDAC1-7) and DNMTs (DNMT1, DNMT3a/b) were supplied by Cell Signaling Technology (Beverly, MA, USA). ..

Article Title: Obesity, Diabetes and Energy Homeostasis: Reduced uterine perfusion pressure causes loss of pancreatic β-cell area but normal function in fetal rat offspring
Article Snippet: .. Placenta or pancreatic lysates were prepared by freezing with liquid nitrogen, processing with mortar and pestle, and then resuspending in 1× radioimmunoprecipitation assay lysis (cat. no. 5871, Cell Signaling) buffer with protease inhibitor cocktail (Cell Signaling). .. Samples were sonicated (Fisher Scientific Sonic Dismembrator model 500; 10 × 3 s, 30% amplitude) and centrifuged to remove the supernatant for total protein quantification via the Pierce BCA Protein Assay Kit (Thermo Scientific), according to the manufacturer’s instructions.

Knock-Out:

Article Title: High Fat Diet Affects Pre-Gestational Adiposity and Glucose Tolerance Perturbing Gestational Placental Macronutrient Transporters Culminating in an Obese Offspring in Wild type and Glucose Transporter Isoform 3 Heterozygous Null Mice
Article Snippet: glut3 +/+ and glut3 +/− embryonic stem cells were cultured in Knock Out DMEM containing high glucose (25 mM) supplemented with 15% FBS and 1% penicillin-streptomycin in a humidified incubator at 37°C in an atmosphere of 5% CO2 and 95% air. .. These cells were then lysed in cell lysis buffer (Cell Signaling Technology, Denvers, MA), supplemented with 1 % phosphatase and protease inhibitor cocktail (Cell Signaling Technology, Danvers, MA).

Concentration Assay:

Article Title: Environmental control of astrocyte pathogenic activities in CNS inflammation
Article Snippet: .. 16 hours later, cells were lysed in ice cold non-denaturing lysis buffer (20 mM Tris-HCl pH=8.0, 137 mM NaCl, 1% NP-40, 2 mM EDTA, recipe obtained from Abcam) supplemented with protease inhibitor cocktail (#5871, Cell Signaling, 1:100 working concentration) and phosphatase inhibitor cocktail (#5870, Cell Signaling, 1:100 working concentration). .. Immunoprecipitation for c-myc was performed for 4 hours at 4C using the following kit (#23620, Thermo Fisher Scientific).

Article Title: High Fat Diet Affects Pre-Gestational Adiposity and Glucose Tolerance Perturbing Gestational Placental Macronutrient Transporters Culminating in an Obese Offspring in Wild type and Glucose Transporter Isoform 3 Heterozygous Null Mice
Article Snippet: These cells were then lysed in cell lysis buffer (Cell Signaling Technology, Denvers, MA), supplemented with 1 % phosphatase and protease inhibitor cocktail (Cell Signaling Technology, Danvers, MA). .. Next glucose uptake experiments were performed under a similar concentration of glucose (2.5 mM) over varying durations.

Lysis:

Article Title: Obesity, Diabetes and Energy Homeostasis: Reduced uterine perfusion pressure causes loss of pancreatic β-cell area but normal function in fetal rat offspring
Article Snippet: .. Placenta or pancreatic lysates were prepared by freezing with liquid nitrogen, processing with mortar and pestle, and then resuspending in 1× radioimmunoprecipitation assay lysis (cat. no. 5871, Cell Signaling) buffer with protease inhibitor cocktail (Cell Signaling). .. Samples were sonicated (Fisher Scientific Sonic Dismembrator model 500; 10 × 3 s, 30% amplitude) and centrifuged to remove the supernatant for total protein quantification via the Pierce BCA Protein Assay Kit (Thermo Scientific), according to the manufacturer’s instructions.

Article Title: Environmental control of astrocyte pathogenic activities in CNS inflammation
Article Snippet: .. 16 hours later, cells were lysed in ice cold non-denaturing lysis buffer (20 mM Tris-HCl pH=8.0, 137 mM NaCl, 1% NP-40, 2 mM EDTA, recipe obtained from Abcam) supplemented with protease inhibitor cocktail (#5871, Cell Signaling, 1:100 working concentration) and phosphatase inhibitor cocktail (#5870, Cell Signaling, 1:100 working concentration). .. Immunoprecipitation for c-myc was performed for 4 hours at 4C using the following kit (#23620, Thermo Fisher Scientific).

Article Title: Novel replisome-associated proteins at cellular replication forks in EBV-transformed B lymphocytes
Article Snippet: After centrifugation, nuclei were resuspended in cold lysis buffer (1% SDS, 50 mM Tris, pH 8.0), sonicated using a microtip sonicator at 8W with 10 sec on and 20 sec off pulses on ice three times to break nuclear membranes. .. Samples were centrifuged at 16,100 × g for 10 min at 4°C to remove debris and diluted with the same volume of ice-cold PBS containing protease inhibitor cocktail (Cell Signaling, #5871), and incubated with pre-washed 150 μl of streptavidin agarose beads (EMD Millipore, #69203) overnight at 4°C.

Article Title: High Fat Diet Affects Pre-Gestational Adiposity and Glucose Tolerance Perturbing Gestational Placental Macronutrient Transporters Culminating in an Obese Offspring in Wild type and Glucose Transporter Isoform 3 Heterozygous Null Mice
Article Snippet: .. These cells were then lysed in cell lysis buffer (Cell Signaling Technology, Denvers, MA), supplemented with 1 % phosphatase and protease inhibitor cocktail (Cell Signaling Technology, Danvers, MA). .. The cell lysates (30 μg) were subjected to Western Blot analysis as described above.

Variant Assay:

Article Title: Environmental control of astrocyte pathogenic activities in CNS inflammation
Article Snippet: HEK293 cells were transfected with human WT IRE1α (a gift from Fumihiko Urano, Addgene plasmid #20744) , Sigmar1-myc-FLAG transcript variant 1 (#RC201206, Origene), or Sigmar1-myc-FLAG transcript variant 2 (#RC214331, Origene) using Lipofectamine 2000 according to the manufacturer’s protocol. .. 16 hours later, cells were lysed in ice cold non-denaturing lysis buffer (20 mM Tris-HCl pH=8.0, 137 mM NaCl, 1% NP-40, 2 mM EDTA, recipe obtained from Abcam) supplemented with protease inhibitor cocktail (#5871, Cell Signaling, 1:100 working concentration) and phosphatase inhibitor cocktail (#5870, Cell Signaling, 1:100 working concentration).

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