radioimmunoprecipitation assay ripa buffer  (Roche)

 
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    Structured Review

    Roche radioimmunoprecipitation assay ripa buffer
    Radioimmunoprecipitation Assay Ripa Buffer, supplied by Roche, used in various techniques. Bioz Stars score: 99/100, based on 454 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/radioimmunoprecipitation assay ripa buffer/product/Roche
    Average 99 stars, based on 454 article reviews
    Price from $9.99 to $1999.99
    radioimmunoprecipitation assay ripa buffer - by Bioz Stars, 2020-04
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    Related Articles

    Centrifugation:

    Article Title: Interferon signaling in Peromyscus leucopus confers a potent and specific restriction to vector-borne flaviviruses
    Article Snippet: Western blotting Cells were washed twice in PBS and lysed in radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl, 150 mM NaCl, 0.1% SDS, 1% NP-40, 0.5% Na-deoxycholate) with complete protease inhibitor cocktail (Roche). .. Cell lysates were treated with Turbo DNase (Thermo Fisher Scientific) and cellular debris was removed by centrifugation (2,000 x g for 5 min) while the supernatant was reserved.

    Article Title: Modulation of histone methylation and MLH1 gene silencing by hexavalent chromium
    Article Snippet: Whole cell lysates were extracted with ice-cold radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl, PH 7.4, 1% NP-40, 0.25% Na-deoxycholate, 150 mM NaCl, 1 mM EDTA) supplemented with protease inhibitor cocktail (Roche Applied Sciences). .. Briefly, cells were lysed with ice-cold RIPA buffer for 10 minutes, followed by centrifugation at 10,000 × g for 10 minutes.

    Article Title: ATR-mediated proteome remodeling is a major determinant of homologous recombination capacity in cancer cells
    Article Snippet: Immunoblotting analysis Cells were harvested and lysed in modified radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris–HCl pH 7.5, 150 mM NaCl, 1% tergitol, 0.25% sodium deoxycholate, 5 mM EDTA) supplemented with Complete EDTA-free protease inhibitor cocktail (Roche), 1 mM PMSF, 5 mM sodium fluoride, 10 mM β-glycerol-phosphate and 0.4 mM sodium orthovanadate. .. Whole cell lysates were cleared with 5 min centrifugation at 13 000 rpm at 4°C and mixed with 3× SDS sample buffer (bromophenol blue, stacking gel buffer, 50% glycerol, 3% SDS and 60 mM DTT).

    Blocking Assay:

    Article Title: Pharmacological activation of TAZ enhances osteogenic differentiation and bone formation of adipose-derived stem cells
    Article Snippet: Western blot analysis and immunoprecipitation assay Cells were lysed with ice-cold radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche). .. After 5% nonfat dry milk or bovine serum albumin (BSA) blocking and overnight incubation with primary antibodies, these blots were detected using appropriate secondary horseradish peroxidase-conjugated secondary antibodies (Invitrogen) and visualized by an enhanced chemiluminescence detection kit (Pierce).

    Real-time Polymerase Chain Reaction:

    Article Title: Transcriptome guided identification of novel functions of RECQ1 helicase
    Article Snippet: For RT-qPCR analysis, 1 µg of total RNA was reverse transcribed using the iScript RT kit (Bio-Rad, catalog no. 1708841), and qPCR was performed using iTaq Universal SYBR Green (Bio-Rad, catalog no. 1725121) as directed by manufacturer. .. To assess the efficiency of knockdown at the protein level, whole-cell lysates were prepared by using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche, catalog no. 04693132001), and the protein concentration was measured using bicinchoninic acid (BCA) protein quantification kit (Thermo Scientific, catalog no. PI23227).

    Incubation:

    Article Title: Pharmacological activation of TAZ enhances osteogenic differentiation and bone formation of adipose-derived stem cells
    Article Snippet: Western blot analysis and immunoprecipitation assay Cells were lysed with ice-cold radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche). .. After 5% nonfat dry milk or bovine serum albumin (BSA) blocking and overnight incubation with primary antibodies, these blots were detected using appropriate secondary horseradish peroxidase-conjugated secondary antibodies (Invitrogen) and visualized by an enhanced chemiluminescence detection kit (Pierce).

    Article Title: Genomics and drug profiling of fatal TCF3-HLF-positive acute lymphoblastic leukemia identifies recurrent mutation patterns and therapeutic options
    Article Snippet: Western blot Whole cell extracts were prepared from 1 × 106 cells using radioimmunoprecipitation assay (RIPA) buffer (20 mM Tris-Cl pH 7.5, 150 mMNaCl, 1% NP-40, 1 mM EDTA pH 8.0, 0.1% SDS) supplemented with Complete mini protease inhibitor cocktail (Roche Life Science) for 20 minutes on ice, sonicated as necessary, and diluted with SDS loading buffer (250 mM Tris pH 6.8, 4% SDS, 0.02% bromophenol blue, 40% glycerol, 4% (v/v) β-mercaptoethanol). .. Membranes were blocked in 5% non-fat dry milk and incubated with primary Bcl-2 (Clone 124; Dako) and tubulin antibodies diluted 1:1,000 in milk.

    Article Title: Parkin deficiency modulates NLRP3 inflammasome activation by attenuating an A20‐dependent negative feedback loop, et al. Parkin deficiency modulates NLRP3 inflammasome activation by attenuating an A20‐dependent negative feedback loop
    Article Snippet: 2.7 Immunoblot analyses and ELISA assays Cell extracts were homogenized in radioimmunoprecipitation assay (RIPA) buffer containing 25 mM β‐glycerophosphate, 50 mM sodium fluoride, 2 mM sodium pyrophosphate, protease inhibitor cocktail (Roche), 1× anti‐phosphatase cocktail mix (100×, Thermo Scientific), and 1× anti‐protease cocktail mix (100×, Thermo Scientific). .. After protein transfer, membranes were blocked in 5% nonfat dry milk in PBS and incubated with primary antibody.

    Article Title: NUAK2 is a critical YAP target in liver cancer
    Article Snippet: Cells and tissue were collected, and processed for western blotting by solubilizing extracts in radioimmunoprecipitation assay (RIPA) buffer with protease inhibitor cocktail (Roche #04693159001) and phosSTOP (Roche, # 04906837001)). .. The gels were blotted onto nitrocellulose or PVDF membranes and blocked in either 5% milk or 1% bovine serum albumin and followed by primary antibody incubation for overnight (NUAK2 (Cat#sc-374348, 1:100, Santa Cruz), YAP (Cat#14071, 1:1000, Cell Signaling), GAPDH (Cat#3683, 1:10000, Cell Signaling), actin (Cat#A5316, 1:1000, Sigma), pMLC (Cat#3671, 1:1000, Cell Signaling), MLC (Cat#M4401, 1:1000, Sigma), TAZ (Cat#4883, 1:1000, Cell Signaling), NF2 (Cat#12888, 1:1000, Cell Signaling)).

    BIA-KA:

    Article Title: Transcriptome guided identification of novel functions of RECQ1 helicase
    Article Snippet: .. To assess the efficiency of knockdown at the protein level, whole-cell lysates were prepared by using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche, catalog no. 04693132001), and the protein concentration was measured using bicinchoninic acid (BCA) protein quantification kit (Thermo Scientific, catalog no. PI23227). .. Ten microgram of total protein per lane was loaded on a SDS-PAGE gel and RECQ1 was detected by immunoblotting using anti-RECQ1 antibody (Santa Cruz, catalog no. SC-25547) at 1:1000 dilution.

    Article Title: Parkin deficiency modulates NLRP3 inflammasome activation by attenuating an A20‐dependent negative feedback loop, et al. Parkin deficiency modulates NLRP3 inflammasome activation by attenuating an A20‐dependent negative feedback loop
    Article Snippet: 2.7 Immunoblot analyses and ELISA assays Cell extracts were homogenized in radioimmunoprecipitation assay (RIPA) buffer containing 25 mM β‐glycerophosphate, 50 mM sodium fluoride, 2 mM sodium pyrophosphate, protease inhibitor cocktail (Roche), 1× anti‐phosphatase cocktail mix (100×, Thermo Scientific), and 1× anti‐protease cocktail mix (100×, Thermo Scientific). .. Protein concentrations were determined with the Micro BCA Protein Assay Reagent Kit (Thermo Scientific), according to the manufacturer's protocol.

    Modification:

    Article Title: ATR-mediated proteome remodeling is a major determinant of homologous recombination capacity in cancer cells
    Article Snippet: .. Immunoblotting analysis Cells were harvested and lysed in modified radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris–HCl pH 7.5, 150 mM NaCl, 1% tergitol, 0.25% sodium deoxycholate, 5 mM EDTA) supplemented with Complete EDTA-free protease inhibitor cocktail (Roche), 1 mM PMSF, 5 mM sodium fluoride, 10 mM β-glycerol-phosphate and 0.4 mM sodium orthovanadate. .. Whole cell lysates were cleared with 5 min centrifugation at 13 000 rpm at 4°C and mixed with 3× SDS sample buffer (bromophenol blue, stacking gel buffer, 50% glycerol, 3% SDS and 60 mM DTT).

    Western Blot:

    Article Title: Interferon signaling in Peromyscus leucopus confers a potent and specific restriction to vector-borne flaviviruses
    Article Snippet: .. Western blotting Cells were washed twice in PBS and lysed in radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl, 150 mM NaCl, 0.1% SDS, 1% NP-40, 0.5% Na-deoxycholate) with complete protease inhibitor cocktail (Roche). .. Cell lysates were treated with Turbo DNase (Thermo Fisher Scientific) and cellular debris was removed by centrifugation (2,000 x g for 5 min) while the supernatant was reserved.

    Article Title: Pharmacological activation of TAZ enhances osteogenic differentiation and bone formation of adipose-derived stem cells
    Article Snippet: .. Western blot analysis and immunoprecipitation assay Cells were lysed with ice-cold radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche). .. Protein lysates were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and then transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore).

    Article Title: Biallelic expansion of an intronic repeat in RFC1 is a common cause of late-onset ataxia
    Article Snippet: Paragraph title: Western blot. ... Cells and tissues were lysed in radioimmunoprecipitation assay (RIPA) buffer supplemented with a complete EDTA-free protease inhibitor cocktail (Roche Diagnostics).

    Article Title: Proliferation of Ewing sarcoma cell lines is suppressed by the receptor tyrosine kinase inhibitors gefitinib and vandetanib
    Article Snippet: Paragraph title: Western blotting ... Following stimulation, cells were scraped off and lysed in radioimmunoprecipitation assay (RIPA) buffer containing 1× Complete Mini Protease Inhibitor (Roche), 50 mM NaF and 1 mM heat-activated Na3 VO4 .

    Article Title: Genomics and drug profiling of fatal TCF3-HLF-positive acute lymphoblastic leukemia identifies recurrent mutation patterns and therapeutic options
    Article Snippet: .. Western blot Whole cell extracts were prepared from 1 × 106 cells using radioimmunoprecipitation assay (RIPA) buffer (20 mM Tris-Cl pH 7.5, 150 mMNaCl, 1% NP-40, 1 mM EDTA pH 8.0, 0.1% SDS) supplemented with Complete mini protease inhibitor cocktail (Roche Life Science) for 20 minutes on ice, sonicated as necessary, and diluted with SDS loading buffer (250 mM Tris pH 6.8, 4% SDS, 0.02% bromophenol blue, 40% glycerol, 4% (v/v) β-mercaptoethanol). .. After SDS-PAGE, proteins were blotted onto nitrocellulose membranes.

    Article Title: NUAK2 is a critical YAP target in liver cancer
    Article Snippet: .. Cells and tissue were collected, and processed for western blotting by solubilizing extracts in radioimmunoprecipitation assay (RIPA) buffer with protease inhibitor cocktail (Roche #04693159001) and phosSTOP (Roche, # 04906837001)). .. Protein lysates were resolved by polyacrylamide gel electrophoresis (PAGE) under reducing conditions (4–12% sodium dodecyl sulfate–PAGE Bis–Tris gels; MOPS buffer system; Invitrogen; NuPAGE-MOPS system).

    Allele-specific Oligonucleotide:

    Article Title: Evaluation of Antisense Oligonucleotides Targeting ATXN3 in SCA3 Mouse Models
    Article Snippet: ASO transfections were performed using Lipofectamine LTX with PLUS Reagent (Life Technologies) at a final ASO concentration of 4 μM/well according to the manufacturer’s instructions. .. Cells were harvested 48 hr after transfection in TRIzol (Invitrogen) for RNA analysis or 72 hr after transfection using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitors (Complete Mini; Roche Diagnostics) for protein analysis.

    Transfection:

    Article Title: Transcriptome guided identification of novel functions of RECQ1 helicase
    Article Snippet: Forty-eight hours after siRNA transfections, total RNA was isolated and whole cell protein lysates were prepared. .. To assess the efficiency of knockdown at the protein level, whole-cell lysates were prepared by using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche, catalog no. 04693132001), and the protein concentration was measured using bicinchoninic acid (BCA) protein quantification kit (Thermo Scientific, catalog no. PI23227).

    Article Title: Evaluation of Antisense Oligonucleotides Targeting ATXN3 in SCA3 Mouse Models
    Article Snippet: .. Cells were harvested 48 hr after transfection in TRIzol (Invitrogen) for RNA analysis or 72 hr after transfection using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitors (Complete Mini; Roche Diagnostics) for protein analysis. .. RNA Isolation and qPCR Total RNA was isolated from transfected fibroblasts and dissected brain tissue using TRIzol reagent according to the manufacturer’s protocol (Invitrogen).

    Concentration Assay:

    Article Title: Evaluation of Antisense Oligonucleotides Targeting ATXN3 in SCA3 Mouse Models
    Article Snippet: ASO transfections were performed using Lipofectamine LTX with PLUS Reagent (Life Technologies) at a final ASO concentration of 4 μM/well according to the manufacturer’s instructions. .. Cells were harvested 48 hr after transfection in TRIzol (Invitrogen) for RNA analysis or 72 hr after transfection using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitors (Complete Mini; Roche Diagnostics) for protein analysis.

    Protease Inhibitor:

    Article Title: Interferon signaling in Peromyscus leucopus confers a potent and specific restriction to vector-borne flaviviruses
    Article Snippet: .. Western blotting Cells were washed twice in PBS and lysed in radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl, 150 mM NaCl, 0.1% SDS, 1% NP-40, 0.5% Na-deoxycholate) with complete protease inhibitor cocktail (Roche). .. Cell lysates were treated with Turbo DNase (Thermo Fisher Scientific) and cellular debris was removed by centrifugation (2,000 x g for 5 min) while the supernatant was reserved.

    Article Title: Pharmacological activation of TAZ enhances osteogenic differentiation and bone formation of adipose-derived stem cells
    Article Snippet: .. Western blot analysis and immunoprecipitation assay Cells were lysed with ice-cold radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche). .. Protein lysates were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and then transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore).

    Article Title: Modulation of histone methylation and MLH1 gene silencing by hexavalent chromium
    Article Snippet: .. Whole cell lysates were extracted with ice-cold radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl, PH 7.4, 1% NP-40, 0.25% Na-deoxycholate, 150 mM NaCl, 1 mM EDTA) supplemented with protease inhibitor cocktail (Roche Applied Sciences). .. Briefly, cells were lysed with ice-cold RIPA buffer for 10 minutes, followed by centrifugation at 10,000 × g for 10 minutes.

    Article Title: Transcriptome guided identification of novel functions of RECQ1 helicase
    Article Snippet: .. To assess the efficiency of knockdown at the protein level, whole-cell lysates were prepared by using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche, catalog no. 04693132001), and the protein concentration was measured using bicinchoninic acid (BCA) protein quantification kit (Thermo Scientific, catalog no. PI23227). .. Ten microgram of total protein per lane was loaded on a SDS-PAGE gel and RECQ1 was detected by immunoblotting using anti-RECQ1 antibody (Santa Cruz, catalog no. SC-25547) at 1:1000 dilution.

    Article Title: Biallelic expansion of an intronic repeat in RFC1 is a common cause of late-onset ataxia
    Article Snippet: .. Cells and tissues were lysed in radioimmunoprecipitation assay (RIPA) buffer supplemented with a complete EDTA-free protease inhibitor cocktail (Roche Diagnostics). .. Brain lysates were homogenized on ice using a tissue ruptor with disposable probes (Qiagen).

    Article Title: Proliferation of Ewing sarcoma cell lines is suppressed by the receptor tyrosine kinase inhibitors gefitinib and vandetanib
    Article Snippet: .. Following stimulation, cells were scraped off and lysed in radioimmunoprecipitation assay (RIPA) buffer containing 1× Complete Mini Protease Inhibitor (Roche), 50 mM NaF and 1 mM heat-activated Na3 VO4 . .. Samples were run on NuPage 4–12% Bis-Tris gels (Invitrogen) using MOPS running buffer and blotted onto PVDF membranes (Immobilon).

    Article Title: Genomics and drug profiling of fatal TCF3-HLF-positive acute lymphoblastic leukemia identifies recurrent mutation patterns and therapeutic options
    Article Snippet: .. Western blot Whole cell extracts were prepared from 1 × 106 cells using radioimmunoprecipitation assay (RIPA) buffer (20 mM Tris-Cl pH 7.5, 150 mMNaCl, 1% NP-40, 1 mM EDTA pH 8.0, 0.1% SDS) supplemented with Complete mini protease inhibitor cocktail (Roche Life Science) for 20 minutes on ice, sonicated as necessary, and diluted with SDS loading buffer (250 mM Tris pH 6.8, 4% SDS, 0.02% bromophenol blue, 40% glycerol, 4% (v/v) β-mercaptoethanol). .. After SDS-PAGE, proteins were blotted onto nitrocellulose membranes.

    Article Title: Parkin deficiency modulates NLRP3 inflammasome activation by attenuating an A20‐dependent negative feedback loop, et al. Parkin deficiency modulates NLRP3 inflammasome activation by attenuating an A20‐dependent negative feedback loop
    Article Snippet: .. 2.7 Immunoblot analyses and ELISA assays Cell extracts were homogenized in radioimmunoprecipitation assay (RIPA) buffer containing 25 mM β‐glycerophosphate, 50 mM sodium fluoride, 2 mM sodium pyrophosphate, protease inhibitor cocktail (Roche), 1× anti‐phosphatase cocktail mix (100×, Thermo Scientific), and 1× anti‐protease cocktail mix (100×, Thermo Scientific). .. Protein concentrations were determined with the Micro BCA Protein Assay Reagent Kit (Thermo Scientific), according to the manufacturer's protocol.

    Article Title: NUAK2 is a critical YAP target in liver cancer
    Article Snippet: .. Cells and tissue were collected, and processed for western blotting by solubilizing extracts in radioimmunoprecipitation assay (RIPA) buffer with protease inhibitor cocktail (Roche #04693159001) and phosSTOP (Roche, # 04906837001)). .. Protein lysates were resolved by polyacrylamide gel electrophoresis (PAGE) under reducing conditions (4–12% sodium dodecyl sulfate–PAGE Bis–Tris gels; MOPS buffer system; Invitrogen; NuPAGE-MOPS system).

    Article Title: ATR-mediated proteome remodeling is a major determinant of homologous recombination capacity in cancer cells
    Article Snippet: .. Immunoblotting analysis Cells were harvested and lysed in modified radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris–HCl pH 7.5, 150 mM NaCl, 1% tergitol, 0.25% sodium deoxycholate, 5 mM EDTA) supplemented with Complete EDTA-free protease inhibitor cocktail (Roche), 1 mM PMSF, 5 mM sodium fluoride, 10 mM β-glycerol-phosphate and 0.4 mM sodium orthovanadate. .. Whole cell lysates were cleared with 5 min centrifugation at 13 000 rpm at 4°C and mixed with 3× SDS sample buffer (bromophenol blue, stacking gel buffer, 50% glycerol, 3% SDS and 60 mM DTT).

    Cell Culture:

    Article Title: Transcriptome guided identification of novel functions of RECQ1 helicase
    Article Snippet: Total RNA from cultured cells was isolated by using Aurum Total RNA Mini Kit (Bio-Rad, catalog no. 7326820) as directed by the manufacturer. .. To assess the efficiency of knockdown at the protein level, whole-cell lysates were prepared by using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche, catalog no. 04693132001), and the protein concentration was measured using bicinchoninic acid (BCA) protein quantification kit (Thermo Scientific, catalog no. PI23227).

    Protein Concentration:

    Article Title: Transcriptome guided identification of novel functions of RECQ1 helicase
    Article Snippet: .. To assess the efficiency of knockdown at the protein level, whole-cell lysates were prepared by using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche, catalog no. 04693132001), and the protein concentration was measured using bicinchoninic acid (BCA) protein quantification kit (Thermo Scientific, catalog no. PI23227). .. Ten microgram of total protein per lane was loaded on a SDS-PAGE gel and RECQ1 was detected by immunoblotting using anti-RECQ1 antibody (Santa Cruz, catalog no. SC-25547) at 1:1000 dilution.

    Sonication:

    Article Title: Genomics and drug profiling of fatal TCF3-HLF-positive acute lymphoblastic leukemia identifies recurrent mutation patterns and therapeutic options
    Article Snippet: .. Western blot Whole cell extracts were prepared from 1 × 106 cells using radioimmunoprecipitation assay (RIPA) buffer (20 mM Tris-Cl pH 7.5, 150 mMNaCl, 1% NP-40, 1 mM EDTA pH 8.0, 0.1% SDS) supplemented with Complete mini protease inhibitor cocktail (Roche Life Science) for 20 minutes on ice, sonicated as necessary, and diluted with SDS loading buffer (250 mM Tris pH 6.8, 4% SDS, 0.02% bromophenol blue, 40% glycerol, 4% (v/v) β-mercaptoethanol). .. After SDS-PAGE, proteins were blotted onto nitrocellulose membranes.

    DC Protein Assay:

    Article Title: Biallelic expansion of an intronic repeat in RFC1 is a common cause of late-onset ataxia
    Article Snippet: Cells and tissues were lysed in radioimmunoprecipitation assay (RIPA) buffer supplemented with a complete EDTA-free protease inhibitor cocktail (Roche Diagnostics). .. Protein lysate concentrations were measured with a DC protein assay (Bio-Rad).

    Nucleic Acid Electrophoresis:

    Article Title: Pharmacological activation of TAZ enhances osteogenic differentiation and bone formation of adipose-derived stem cells
    Article Snippet: Western blot analysis and immunoprecipitation assay Cells were lysed with ice-cold radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche). .. Protein lysates were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and then transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore).

    Isolation:

    Article Title: Transcriptome guided identification of novel functions of RECQ1 helicase
    Article Snippet: Total RNA from cultured cells was isolated by using Aurum Total RNA Mini Kit (Bio-Rad, catalog no. 7326820) as directed by the manufacturer. .. To assess the efficiency of knockdown at the protein level, whole-cell lysates were prepared by using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche, catalog no. 04693132001), and the protein concentration was measured using bicinchoninic acid (BCA) protein quantification kit (Thermo Scientific, catalog no. PI23227).

    Article Title: The endocrine disruptor DEHP and the ECS: analysis of a possible crosstalk
    Article Snippet: .. Protein was isolated using radioimmunoprecipitation assay (RIPA) buffer including protease and phosphatase inhibitors (Roche). .. Protein concentration was determined by the BioRad Protein Assay (BioRad).

    Hormonal Assay:

    Article Title: The endocrine disruptor DEHP and the ECS: analysis of a possible crosstalk
    Article Snippet: Paragraph title: Hormone assay ... Protein was isolated using radioimmunoprecipitation assay (RIPA) buffer including protease and phosphatase inhibitors (Roche).

    Quantitative RT-PCR:

    Article Title: Transcriptome guided identification of novel functions of RECQ1 helicase
    Article Snippet: Paragraph title: 2.2. Validation of knockdown by RT-qPCR and immunoblotting ... To assess the efficiency of knockdown at the protein level, whole-cell lysates were prepared by using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche, catalog no. 04693132001), and the protein concentration was measured using bicinchoninic acid (BCA) protein quantification kit (Thermo Scientific, catalog no. PI23227).

    Polyacrylamide Gel Electrophoresis:

    Article Title: NUAK2 is a critical YAP target in liver cancer
    Article Snippet: Cells and tissue were collected, and processed for western blotting by solubilizing extracts in radioimmunoprecipitation assay (RIPA) buffer with protease inhibitor cocktail (Roche #04693159001) and phosSTOP (Roche, # 04906837001)). .. Protein lysates were resolved by polyacrylamide gel electrophoresis (PAGE) under reducing conditions (4–12% sodium dodecyl sulfate–PAGE Bis–Tris gels; MOPS buffer system; Invitrogen; NuPAGE-MOPS system).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: NUAK2 is a critical YAP target in liver cancer
    Article Snippet: Cells and tissue were collected, and processed for western blotting by solubilizing extracts in radioimmunoprecipitation assay (RIPA) buffer with protease inhibitor cocktail (Roche #04693159001) and phosSTOP (Roche, # 04906837001)). .. The gels were blotted onto nitrocellulose or PVDF membranes and blocked in either 5% milk or 1% bovine serum albumin and followed by primary antibody incubation for overnight (NUAK2 (Catsc-374348, 1:100, Santa Cruz), YAP (Cat#14071, 1:1000, Cell Signaling), GAPDH (Cat#3683, 1:10000, Cell Signaling), actin (Cat#A5316, 1:1000, Sigma), pMLC (Cat#3671, 1:1000, Cell Signaling), MLC (Cat#M4401, 1:1000, Sigma), TAZ (Cat#4883, 1:1000, Cell Signaling), NF2 (Cat#12888, 1:1000, Cell Signaling)).

    SDS Page:

    Article Title: Pharmacological activation of TAZ enhances osteogenic differentiation and bone formation of adipose-derived stem cells
    Article Snippet: Western blot analysis and immunoprecipitation assay Cells were lysed with ice-cold radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche). .. Protein lysates were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and then transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore).

    Article Title: Transcriptome guided identification of novel functions of RECQ1 helicase
    Article Snippet: To assess the efficiency of knockdown at the protein level, whole-cell lysates were prepared by using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche, catalog no. 04693132001), and the protein concentration was measured using bicinchoninic acid (BCA) protein quantification kit (Thermo Scientific, catalog no. PI23227). .. Ten microgram of total protein per lane was loaded on a SDS-PAGE gel and RECQ1 was detected by immunoblotting using anti-RECQ1 antibody (Santa Cruz, catalog no. SC-25547) at 1:1000 dilution.

    Article Title: Genomics and drug profiling of fatal TCF3-HLF-positive acute lymphoblastic leukemia identifies recurrent mutation patterns and therapeutic options
    Article Snippet: Western blot Whole cell extracts were prepared from 1 × 106 cells using radioimmunoprecipitation assay (RIPA) buffer (20 mM Tris-Cl pH 7.5, 150 mMNaCl, 1% NP-40, 1 mM EDTA pH 8.0, 0.1% SDS) supplemented with Complete mini protease inhibitor cocktail (Roche Life Science) for 20 minutes on ice, sonicated as necessary, and diluted with SDS loading buffer (250 mM Tris pH 6.8, 4% SDS, 0.02% bromophenol blue, 40% glycerol, 4% (v/v) β-mercaptoethanol). .. After SDS-PAGE, proteins were blotted onto nitrocellulose membranes.

    Article Title: ATR-mediated proteome remodeling is a major determinant of homologous recombination capacity in cancer cells
    Article Snippet: Immunoblotting analysis Cells were harvested and lysed in modified radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris–HCl pH 7.5, 150 mM NaCl, 1% tergitol, 0.25% sodium deoxycholate, 5 mM EDTA) supplemented with Complete EDTA-free protease inhibitor cocktail (Roche), 1 mM PMSF, 5 mM sodium fluoride, 10 mM β-glycerol-phosphate and 0.4 mM sodium orthovanadate. .. After resolved on SDS-PAGE gels, proteins were then transferred onto polyvinylidene difluoride (PVDF) membranes and detected with designated antibodies.

    SYBR Green Assay:

    Article Title: Transcriptome guided identification of novel functions of RECQ1 helicase
    Article Snippet: For RT-qPCR analysis, 1 µg of total RNA was reverse transcribed using the iScript RT kit (Bio-Rad, catalog no. 1708841), and qPCR was performed using iTaq Universal SYBR Green (Bio-Rad, catalog no. 1725121) as directed by manufacturer. .. To assess the efficiency of knockdown at the protein level, whole-cell lysates were prepared by using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche, catalog no. 04693132001), and the protein concentration was measured using bicinchoninic acid (BCA) protein quantification kit (Thermo Scientific, catalog no. PI23227).

    Radio Immunoprecipitation:

    Article Title: Interferon signaling in Peromyscus leucopus confers a potent and specific restriction to vector-borne flaviviruses
    Article Snippet: .. Western blotting Cells were washed twice in PBS and lysed in radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl, 150 mM NaCl, 0.1% SDS, 1% NP-40, 0.5% Na-deoxycholate) with complete protease inhibitor cocktail (Roche). .. Cell lysates were treated with Turbo DNase (Thermo Fisher Scientific) and cellular debris was removed by centrifugation (2,000 x g for 5 min) while the supernatant was reserved.

    Article Title: Pharmacological activation of TAZ enhances osteogenic differentiation and bone formation of adipose-derived stem cells
    Article Snippet: .. Western blot analysis and immunoprecipitation assay Cells were lysed with ice-cold radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche). .. Protein lysates were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and then transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore).

    Article Title: Modulation of histone methylation and MLH1 gene silencing by hexavalent chromium
    Article Snippet: .. Whole cell lysates were extracted with ice-cold radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl, PH 7.4, 1% NP-40, 0.25% Na-deoxycholate, 150 mM NaCl, 1 mM EDTA) supplemented with protease inhibitor cocktail (Roche Applied Sciences). .. Briefly, cells were lysed with ice-cold RIPA buffer for 10 minutes, followed by centrifugation at 10,000 × g for 10 minutes.

    Article Title: Transcriptome guided identification of novel functions of RECQ1 helicase
    Article Snippet: .. To assess the efficiency of knockdown at the protein level, whole-cell lysates were prepared by using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche, catalog no. 04693132001), and the protein concentration was measured using bicinchoninic acid (BCA) protein quantification kit (Thermo Scientific, catalog no. PI23227). .. Ten microgram of total protein per lane was loaded on a SDS-PAGE gel and RECQ1 was detected by immunoblotting using anti-RECQ1 antibody (Santa Cruz, catalog no. SC-25547) at 1:1000 dilution.

    Article Title: Evaluation of Antisense Oligonucleotides Targeting ATXN3 in SCA3 Mouse Models
    Article Snippet: .. Cells were harvested 48 hr after transfection in TRIzol (Invitrogen) for RNA analysis or 72 hr after transfection using radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitors (Complete Mini; Roche Diagnostics) for protein analysis. .. RNA Isolation and qPCR Total RNA was isolated from transfected fibroblasts and dissected brain tissue using TRIzol reagent according to the manufacturer’s protocol (Invitrogen).

    Article Title: Biallelic expansion of an intronic repeat in RFC1 is a common cause of late-onset ataxia
    Article Snippet: .. Cells and tissues were lysed in radioimmunoprecipitation assay (RIPA) buffer supplemented with a complete EDTA-free protease inhibitor cocktail (Roche Diagnostics). .. Brain lysates were homogenized on ice using a tissue ruptor with disposable probes (Qiagen).

    Article Title: Proliferation of Ewing sarcoma cell lines is suppressed by the receptor tyrosine kinase inhibitors gefitinib and vandetanib
    Article Snippet: .. Following stimulation, cells were scraped off and lysed in radioimmunoprecipitation assay (RIPA) buffer containing 1× Complete Mini Protease Inhibitor (Roche), 50 mM NaF and 1 mM heat-activated Na3 VO4 . .. Samples were run on NuPage 4–12% Bis-Tris gels (Invitrogen) using MOPS running buffer and blotted onto PVDF membranes (Immobilon).

    Article Title: Genomics and drug profiling of fatal TCF3-HLF-positive acute lymphoblastic leukemia identifies recurrent mutation patterns and therapeutic options
    Article Snippet: .. Western blot Whole cell extracts were prepared from 1 × 106 cells using radioimmunoprecipitation assay (RIPA) buffer (20 mM Tris-Cl pH 7.5, 150 mMNaCl, 1% NP-40, 1 mM EDTA pH 8.0, 0.1% SDS) supplemented with Complete mini protease inhibitor cocktail (Roche Life Science) for 20 minutes on ice, sonicated as necessary, and diluted with SDS loading buffer (250 mM Tris pH 6.8, 4% SDS, 0.02% bromophenol blue, 40% glycerol, 4% (v/v) β-mercaptoethanol). .. After SDS-PAGE, proteins were blotted onto nitrocellulose membranes.

    Article Title: Parkin deficiency modulates NLRP3 inflammasome activation by attenuating an A20‐dependent negative feedback loop, et al. Parkin deficiency modulates NLRP3 inflammasome activation by attenuating an A20‐dependent negative feedback loop
    Article Snippet: .. 2.7 Immunoblot analyses and ELISA assays Cell extracts were homogenized in radioimmunoprecipitation assay (RIPA) buffer containing 25 mM β‐glycerophosphate, 50 mM sodium fluoride, 2 mM sodium pyrophosphate, protease inhibitor cocktail (Roche), 1× anti‐phosphatase cocktail mix (100×, Thermo Scientific), and 1× anti‐protease cocktail mix (100×, Thermo Scientific). .. Protein concentrations were determined with the Micro BCA Protein Assay Reagent Kit (Thermo Scientific), according to the manufacturer's protocol.

    Article Title: NUAK2 is a critical YAP target in liver cancer
    Article Snippet: .. Cells and tissue were collected, and processed for western blotting by solubilizing extracts in radioimmunoprecipitation assay (RIPA) buffer with protease inhibitor cocktail (Roche #04693159001) and phosSTOP (Roche, # 04906837001)). .. Protein lysates were resolved by polyacrylamide gel electrophoresis (PAGE) under reducing conditions (4–12% sodium dodecyl sulfate–PAGE Bis–Tris gels; MOPS buffer system; Invitrogen; NuPAGE-MOPS system).

    Article Title: The endocrine disruptor DEHP and the ECS: analysis of a possible crosstalk
    Article Snippet: .. Protein was isolated using radioimmunoprecipitation assay (RIPA) buffer including protease and phosphatase inhibitors (Roche). .. Protein concentration was determined by the BioRad Protein Assay (BioRad).

    Article Title: ATR-mediated proteome remodeling is a major determinant of homologous recombination capacity in cancer cells
    Article Snippet: .. Immunoblotting analysis Cells were harvested and lysed in modified radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris–HCl pH 7.5, 150 mM NaCl, 1% tergitol, 0.25% sodium deoxycholate, 5 mM EDTA) supplemented with Complete EDTA-free protease inhibitor cocktail (Roche), 1 mM PMSF, 5 mM sodium fluoride, 10 mM β-glycerol-phosphate and 0.4 mM sodium orthovanadate. .. Whole cell lysates were cleared with 5 min centrifugation at 13 000 rpm at 4°C and mixed with 3× SDS sample buffer (bromophenol blue, stacking gel buffer, 50% glycerol, 3% SDS and 60 mM DTT).

    Electrophoresis:

    Article Title: Interferon signaling in Peromyscus leucopus confers a potent and specific restriction to vector-borne flaviviruses
    Article Snippet: Western blotting Cells were washed twice in PBS and lysed in radioimmunoprecipitation assay (RIPA) buffer (50 mM Tris-HCl, 150 mM NaCl, 0.1% SDS, 1% NP-40, 0.5% Na-deoxycholate) with complete protease inhibitor cocktail (Roche). .. Equal amounts (10–30 μg) of protein were loaded on a 10% polyacrylamide gel and resolved by electrophoresis.

    Enzyme-linked Immunosorbent Assay:

    Article Title: Parkin deficiency modulates NLRP3 inflammasome activation by attenuating an A20‐dependent negative feedback loop, et al. Parkin deficiency modulates NLRP3 inflammasome activation by attenuating an A20‐dependent negative feedback loop
    Article Snippet: .. 2.7 Immunoblot analyses and ELISA assays Cell extracts were homogenized in radioimmunoprecipitation assay (RIPA) buffer containing 25 mM β‐glycerophosphate, 50 mM sodium fluoride, 2 mM sodium pyrophosphate, protease inhibitor cocktail (Roche), 1× anti‐phosphatase cocktail mix (100×, Thermo Scientific), and 1× anti‐protease cocktail mix (100×, Thermo Scientific). .. Protein concentrations were determined with the Micro BCA Protein Assay Reagent Kit (Thermo Scientific), according to the manufacturer's protocol.

    Immunoprecipitation:

    Article Title: Pharmacological activation of TAZ enhances osteogenic differentiation and bone formation of adipose-derived stem cells
    Article Snippet: .. Western blot analysis and immunoprecipitation assay Cells were lysed with ice-cold radioimmunoprecipitation assay (RIPA) buffer containing protease inhibitor cocktail (Roche). .. Protein lysates were resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and then transferred onto polyvinylidene difluoride (PVDF) membranes (Millipore).

    Staining:

    Article Title: Proliferation of Ewing sarcoma cell lines is suppressed by the receptor tyrosine kinase inhibitors gefitinib and vandetanib
    Article Snippet: Following stimulation, cells were scraped off and lysed in radioimmunoprecipitation assay (RIPA) buffer containing 1× Complete Mini Protease Inhibitor (Roche), 50 mM NaF and 1 mM heat-activated Na3 VO4 . .. Blots were stained with antibodies for p44/42 MAPK (137F5, Cell Signaling Technology), phospho-p44/42 MAPK (20G11, Cell Signaling Technology), Akt1 (2H10, Cell Signaling Technology), phospho-Akt (587F11, Cell Signaling Technology), cyclin D1 (M7155, DakoCytomation), c-Myc (sc-40, Santa Cruz Biotechnology) and β-actin (sc-8432, Santa Cruz Biotechnology).

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    Roche radioimmunoprecipitaion assay buffer
    Radioimmunoprecipitaion Assay Buffer, supplied by Roche, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    radioimmunoprecipitaion assay buffer - by Bioz Stars, 2020-04
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