lysis buffer  (Thermo Fisher)


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    Name:
    Lysis Buffer A
    Description:
    Thermo Scientific GeneJET Lysis Buffer A is a component of the GeneJET Plant Genomic DNAPurification Mini Kit K0791 K0792 and may be purchased separately
    Catalog Number:
    r1661
    Price:
    None
    Applications:
    DNA & RNA Purification & Analysis|DNA Extraction|Genomic DNA Purification|gDNA from Plant & Fungi
    Category:
    Lab Reagents and Chemicals
    Buy from Supplier


    Structured Review

    Thermo Fisher lysis buffer
    Thermo Scientific GeneJET Lysis Buffer A is a component of the GeneJET Plant Genomic DNAPurification Mini Kit K0791 K0792 and may be purchased separately
    https://www.bioz.com/result/lysis buffer/product/Thermo Fisher
    Average 99 stars, based on 2857 article reviews
    Price from $9.99 to $1999.99
    lysis buffer - by Bioz Stars, 2020-09
    99/100 stars

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    Related Articles

    Sonication:

    Article Title: The WblC/WhiB7 Transcription Factor Controls Intrinsic Resistance to Translation-Targeting Antibiotics by Altering Ribosome Composition
    Article Snippet: .. Then, the cells were harvested and washed with wash buffer (20 mM Tris-Cl [pH 7.4], 100 mM NaCl, 10 mM MgCl2 ) and were lysed by sonication in lysis buffer (20 mM Tris-Cl [pH 7.4], 200 mM NH4 Cl, 10 mM MgCl2 , 0.5 mM EDTA, 6 mM β-mercaptoethanol) with 5 U/ml RNase inhibitor (Applied Biosystems) and 1 mM phenylmethylsulfonyl fluoride (PMSF). .. After 5 U/ml Turbo DNase (Ambion) treatment at 4°C for 15 min, lysates were clarified twice by centrifugation (22,000 × g, 10 min, 4°C).

    Proximity Ligation Assay:

    Article Title: Identification of inhibitors regulating cell proliferation and FUS-DDIT3 expression in myxoid liposarcoma using combined DNA, mRNA, and protein analyses
    Article Snippet: .. Therefore, to test how efficiently we could quantify FUS-DDIT3 and FUS we also tested a standard PLA lysis buffer (Protein Quant Sample Lysis Buffer, Applied Biosystems). .. Supplementary Figure shows that the BSA direct lysis buffer is superior the PLA lysis buffer to quantify DNA, while they were equally suitable for quantifying mRNA.

    Isolation:

    Article Title: Kidney micro-organoids in suspension culture as a scalable source of human pluripotent stem cell-derived kidney cells
    Article Snippet: .. Real time-qPCR analysis Organoids were lysed in lysis buffer and mRNA was isolated from organoids using the Ambion mRNA Isolation Kit (Life Technologies) according to the manufacturer's instructions. .. We converted 0.5-1 µg mRNA to cDNA in the presence of thermostable RNAse inhibitor and a reaction mix containing GoScript reverse transcriptase (Promega), MgCl2 , nucleotide mix and 5× reaction buffers, using standard GoScript reverse transcription protocol.

    Article Title: Neutrophils prime a long-lived effector macrophage phenotype that mediates accelerated helminth expulsion
    Article Snippet: .. Gene expression by microarray and qPCR For microarray, lung macrophages (10,000 cells) were directly sorted into RNA lysis buffer (Ambion RNAqueous Micro for RNA isolation) and frozen on dry ice. .. RNA was isolated, amplified using the AminoAllyl MessageAmp II kit (Life Technologies, Grand Island, NY), coupled to Cy3, and hybridized to SurePrint Mouse 8x60k gene expression microarrays (Agilent Technologies, Santa Clara CA) as per manufacturer recommendations.

    Article Title: The long non-coding RNA Meg3 is dispensable for hematopoietic stem cells
    Article Snippet: .. Gene Expression Analysis by qPCR HSCs (MxCre or MxCre Meg3mat flox/pat wt LSK CD150+ CD48− CD34− or fetal liver HSCs: embryonic stage 13.5 CD48− CD41+ cKit+ CD34+; adult HSCs (8–12 weeks): LSK CD150+ CD48− CD34− CD135−; aged HSCs (24 month): LSK CD150+ CD48− CD34− CD135−) were FACS sorted into RNA lysis buffer (ARCTURUS PicoPure RNA Isolation Kit (Life Technologies, Invitrogen, Carlsbad, CA, USA)) for qPCR analysis and stored at −80 °C. .. RNA was isolated using the ARCTURUS PicoPure RNA Isolation Kit (Life Technologies, Invitrogen, Carlsbad, CA, USA) following manufacturer’s instructions.

    Real-time Polymerase Chain Reaction:

    Article Title: Neutrophils prime a long-lived effector macrophage phenotype that mediates accelerated helminth expulsion
    Article Snippet: .. Gene expression by microarray and qPCR For microarray, lung macrophages (10,000 cells) were directly sorted into RNA lysis buffer (Ambion RNAqueous Micro for RNA isolation) and frozen on dry ice. .. RNA was isolated, amplified using the AminoAllyl MessageAmp II kit (Life Technologies, Grand Island, NY), coupled to Cy3, and hybridized to SurePrint Mouse 8x60k gene expression microarrays (Agilent Technologies, Santa Clara CA) as per manufacturer recommendations.

    Article Title: The long non-coding RNA Meg3 is dispensable for hematopoietic stem cells
    Article Snippet: .. Gene Expression Analysis by qPCR HSCs (MxCre or MxCre Meg3mat flox/pat wt LSK CD150+ CD48− CD34− or fetal liver HSCs: embryonic stage 13.5 CD48− CD41+ cKit+ CD34+; adult HSCs (8–12 weeks): LSK CD150+ CD48− CD34− CD135−; aged HSCs (24 month): LSK CD150+ CD48− CD34− CD135−) were FACS sorted into RNA lysis buffer (ARCTURUS PicoPure RNA Isolation Kit (Life Technologies, Invitrogen, Carlsbad, CA, USA)) for qPCR analysis and stored at −80 °C. .. RNA was isolated using the ARCTURUS PicoPure RNA Isolation Kit (Life Technologies, Invitrogen, Carlsbad, CA, USA) following manufacturer’s instructions.

    Microarray:

    Article Title: Neutrophils prime a long-lived effector macrophage phenotype that mediates accelerated helminth expulsion
    Article Snippet: .. Gene expression by microarray and qPCR For microarray, lung macrophages (10,000 cells) were directly sorted into RNA lysis buffer (Ambion RNAqueous Micro for RNA isolation) and frozen on dry ice. .. RNA was isolated, amplified using the AminoAllyl MessageAmp II kit (Life Technologies, Grand Island, NY), coupled to Cy3, and hybridized to SurePrint Mouse 8x60k gene expression microarrays (Agilent Technologies, Santa Clara CA) as per manufacturer recommendations.

    Incubation:

    Article Title: Functional Mapping of Transcription Factor Grf10 That Regulates Adenine-Responsive and Filamentation Genes in Candida albicans
    Article Snippet: .. Briefly, the frozen cell pellet was resuspended in 200 µl of lysis buffer (0.1 M NaOH, 0.5 M EDTA, 2% SDS, 2% β-mercaptoethanol, protease inhibitor cocktail mix [Thermo Scientific]), and incubated for 10 min at 90°C. ..

    Expressing:

    Article Title: Neutrophils prime a long-lived effector macrophage phenotype that mediates accelerated helminth expulsion
    Article Snippet: .. Gene expression by microarray and qPCR For microarray, lung macrophages (10,000 cells) were directly sorted into RNA lysis buffer (Ambion RNAqueous Micro for RNA isolation) and frozen on dry ice. .. RNA was isolated, amplified using the AminoAllyl MessageAmp II kit (Life Technologies, Grand Island, NY), coupled to Cy3, and hybridized to SurePrint Mouse 8x60k gene expression microarrays (Agilent Technologies, Santa Clara CA) as per manufacturer recommendations.

    Article Title: The long non-coding RNA Meg3 is dispensable for hematopoietic stem cells
    Article Snippet: .. Gene Expression Analysis by qPCR HSCs (MxCre or MxCre Meg3mat flox/pat wt LSK CD150+ CD48− CD34− or fetal liver HSCs: embryonic stage 13.5 CD48− CD41+ cKit+ CD34+; adult HSCs (8–12 weeks): LSK CD150+ CD48− CD34− CD135−; aged HSCs (24 month): LSK CD150+ CD48− CD34− CD135−) were FACS sorted into RNA lysis buffer (ARCTURUS PicoPure RNA Isolation Kit (Life Technologies, Invitrogen, Carlsbad, CA, USA)) for qPCR analysis and stored at −80 °C. .. RNA was isolated using the ARCTURUS PicoPure RNA Isolation Kit (Life Technologies, Invitrogen, Carlsbad, CA, USA) following manufacturer’s instructions.

    Protease Inhibitor:

    Article Title: Functional Mapping of Transcription Factor Grf10 That Regulates Adenine-Responsive and Filamentation Genes in Candida albicans
    Article Snippet: .. Briefly, the frozen cell pellet was resuspended in 200 µl of lysis buffer (0.1 M NaOH, 0.5 M EDTA, 2% SDS, 2% β-mercaptoethanol, protease inhibitor cocktail mix [Thermo Scientific]), and incubated for 10 min at 90°C. ..

    FACS:

    Article Title: The long non-coding RNA Meg3 is dispensable for hematopoietic stem cells
    Article Snippet: .. Gene Expression Analysis by qPCR HSCs (MxCre or MxCre Meg3mat flox/pat wt LSK CD150+ CD48− CD34− or fetal liver HSCs: embryonic stage 13.5 CD48− CD41+ cKit+ CD34+; adult HSCs (8–12 weeks): LSK CD150+ CD48− CD34− CD135−; aged HSCs (24 month): LSK CD150+ CD48− CD34− CD135−) were FACS sorted into RNA lysis buffer (ARCTURUS PicoPure RNA Isolation Kit (Life Technologies, Invitrogen, Carlsbad, CA, USA)) for qPCR analysis and stored at −80 °C. .. RNA was isolated using the ARCTURUS PicoPure RNA Isolation Kit (Life Technologies, Invitrogen, Carlsbad, CA, USA) following manufacturer’s instructions.

    Lysis:

    Article Title: Kidney micro-organoids in suspension culture as a scalable source of human pluripotent stem cell-derived kidney cells
    Article Snippet: .. Real time-qPCR analysis Organoids were lysed in lysis buffer and mRNA was isolated from organoids using the Ambion mRNA Isolation Kit (Life Technologies) according to the manufacturer's instructions. .. We converted 0.5-1 µg mRNA to cDNA in the presence of thermostable RNAse inhibitor and a reaction mix containing GoScript reverse transcriptase (Promega), MgCl2 , nucleotide mix and 5× reaction buffers, using standard GoScript reverse transcription protocol.

    Article Title: Neutrophils prime a long-lived effector macrophage phenotype that mediates accelerated helminth expulsion
    Article Snippet: .. Gene expression by microarray and qPCR For microarray, lung macrophages (10,000 cells) were directly sorted into RNA lysis buffer (Ambion RNAqueous Micro for RNA isolation) and frozen on dry ice. .. RNA was isolated, amplified using the AminoAllyl MessageAmp II kit (Life Technologies, Grand Island, NY), coupled to Cy3, and hybridized to SurePrint Mouse 8x60k gene expression microarrays (Agilent Technologies, Santa Clara CA) as per manufacturer recommendations.

    Article Title: Characterisation of immune cell function in fragment and full-length Huntington's disease mouse models
    Article Snippet: .. Blood monocytes Blood was obtained by cardiac puncture, collected into EDTA coated tubes (BD Vacutainer), and 1 ml red blood cell lysis buffer (ammonium chloride buffer; eBioscience) was added per 1 ml of blood. .. Following centrifugation at 300 × g for 5 min, the lysis step was repeated twice.

    Article Title: The WblC/WhiB7 Transcription Factor Controls Intrinsic Resistance to Translation-Targeting Antibiotics by Altering Ribosome Composition
    Article Snippet: .. Then, the cells were harvested and washed with wash buffer (20 mM Tris-Cl [pH 7.4], 100 mM NaCl, 10 mM MgCl2 ) and were lysed by sonication in lysis buffer (20 mM Tris-Cl [pH 7.4], 200 mM NH4 Cl, 10 mM MgCl2 , 0.5 mM EDTA, 6 mM β-mercaptoethanol) with 5 U/ml RNase inhibitor (Applied Biosystems) and 1 mM phenylmethylsulfonyl fluoride (PMSF). .. After 5 U/ml Turbo DNase (Ambion) treatment at 4°C for 15 min, lysates were clarified twice by centrifugation (22,000 × g, 10 min, 4°C).

    Article Title: Functional Mapping of Transcription Factor Grf10 That Regulates Adenine-Responsive and Filamentation Genes in Candida albicans
    Article Snippet: .. Briefly, the frozen cell pellet was resuspended in 200 µl of lysis buffer (0.1 M NaOH, 0.5 M EDTA, 2% SDS, 2% β-mercaptoethanol, protease inhibitor cocktail mix [Thermo Scientific]), and incubated for 10 min at 90°C. ..

    Article Title: The long non-coding RNA Meg3 is dispensable for hematopoietic stem cells
    Article Snippet: .. Gene Expression Analysis by qPCR HSCs (MxCre or MxCre Meg3mat flox/pat wt LSK CD150+ CD48− CD34− or fetal liver HSCs: embryonic stage 13.5 CD48− CD41+ cKit+ CD34+; adult HSCs (8–12 weeks): LSK CD150+ CD48− CD34− CD135−; aged HSCs (24 month): LSK CD150+ CD48− CD34− CD135−) were FACS sorted into RNA lysis buffer (ARCTURUS PicoPure RNA Isolation Kit (Life Technologies, Invitrogen, Carlsbad, CA, USA)) for qPCR analysis and stored at −80 °C. .. RNA was isolated using the ARCTURUS PicoPure RNA Isolation Kit (Life Technologies, Invitrogen, Carlsbad, CA, USA) following manufacturer’s instructions.

    Article Title: Identification of inhibitors regulating cell proliferation and FUS-DDIT3 expression in myxoid liposarcoma using combined DNA, mRNA, and protein analyses
    Article Snippet: .. Therefore, to test how efficiently we could quantify FUS-DDIT3 and FUS we also tested a standard PLA lysis buffer (Protein Quant Sample Lysis Buffer, Applied Biosystems). .. Supplementary Figure shows that the BSA direct lysis buffer is superior the PLA lysis buffer to quantify DNA, while they were equally suitable for quantifying mRNA.

    Article Title: H3K27me3 demethylases regulate in vitro chondrogenesis and chondrocyte activity in osteoarthritis
    Article Snippet: .. Treated HACs and MSCs were washed twice in PBS, and cells were harvested in cells to complementary DNA (cDNA) lysis buffer (Ambion; Thermo Fisher Scientific, Austin, TX, USA) prior to cDNA synthesis. ..

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  • 99
    Thermo Fisher sybr green i nucleic acid gel stain 10
    Effect of synchrony on drug susceptibility measured across the life cycle using the <t>SYBR</t> <t>Green</t> I method after one or two life cycles. EC 50 values for chloroquine (squares), dihydroartemisinin (circles) or pyrimethamine (diamonds) were determined from experiments initiated at the times shown on the y -axis using synchronized parasites at 1% parasitemia and 1% haematocrit that were incubated for either 27 h (a) or 54 h (b) for P. knowlesi , and 48 h (c) or 96 h (d) for P. falciparum . Signal window (squares) and assay quality ( Z ′ factor, circles) (e–h) were determined as in Figure 1 .
    Sybr Green I Nucleic Acid Gel Stain 10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr green i nucleic acid gel stain 10/product/Thermo Fisher
    Average 99 stars, based on 26 article reviews
    Price from $9.99 to $1999.99
    sybr green i nucleic acid gel stain 10 - by Bioz Stars, 2020-09
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    84
    Thermo Fisher ice cold immunoprecipitation lysis buffer
    ARRB1 interacts with ASK1 and then antagonizes its TRAF6‐mediated K6‐linked polyubiquitination, thus inhibiting its activation in hepatocytes during hepatic I/R injury. A, Immunoblotting detection of phosphorylated ASK1 protein (left panel) and co‐IP assay for the interaction between His‐ASK1 and HA‐TRAF proteins (right panel) in L02 hepatocytes stimulated with H/R for 6 h. B, His‐tagged ASK1 and HA‐tagged TRAF6 plasmids were cotransfected into L02 hepatocytes. Anti‐His antibody (left panel) and anti‐HA antibody (right panel) were used for <t>immunoprecipitation.</t> C, Immunoprecipitation analysis of indicated ubiquitination types of ASK1 in L02 hepatocytes transfected with HA‐TRAF6 or empty vector in respond to H/R insult or not. D, IP analysis was conducted to detect the binding association between ASK1 and ARRB1 in L02 hepatocytes under normal condition. His‐tagged ASK1 and Flag‐tagged ARRB1 plasmids were cotransfected into L02 hepatocytes. Anti‐His antibody (left panel) and anti‐Flag antibody (right panel) were used for immunoprecipitation. E, IP analysis was conducted to detect the binding association between ASK1 and ARRB1 in L02 hepatocytes when treated with H/R challenge. Anti‐His antibody (left panel) and anti‐Flag antibody (right panel) were used for immunoprecipitation. F, IP analysis showing the expression level of ARRB1 and binding capability between ASK1 and TRAF6 in L02 hepatocytes under H/R challenge or not. Anti‐His antibody (left panel) and anti‐HA antibody (right panel) were used for immunoprecipitation. G and H, Lysates of liver lobes challenged with or without I/R surgery (G) or whole‐cell lysates of primary hepatocytes subjected to or not H/R insult (H) were subjected to immunoprecipitation with anti‐ASK1 antibody followed by immunoblotting with anti‐K6‐linked polyubiquitination antibody when ARRB1 was overexpressed or knockout
    Ice Cold Immunoprecipitation Lysis Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ice cold immunoprecipitation lysis buffer/product/Thermo Fisher
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ice cold immunoprecipitation lysis buffer - by Bioz Stars, 2020-09
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    99
    Thermo Fisher ripa buffer lyse
    Time curve showing mtDNA content measured in a blood sample extracted by BioRobot EZ1 Advanced after incubation using 4 different lysis buffers. The buffers used were <t>G2</t> (included in the EZ1 Tissue kit) and Pierce <t>RIPA</t> Buffer Lyse (Thermo Fisher Scientific, Waltham, MA, USA). Both buffers were used pure and in combination with Proteinase K (PK). The baseline is obtained using the standard EZ1 tissue kit protocol without incubation.
    Ripa Buffer Lyse, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ripa buffer lyse/product/Thermo Fisher
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    ripa buffer lyse - by Bioz Stars, 2020-09
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    Image Search Results


    Effect of synchrony on drug susceptibility measured across the life cycle using the SYBR Green I method after one or two life cycles. EC 50 values for chloroquine (squares), dihydroartemisinin (circles) or pyrimethamine (diamonds) were determined from experiments initiated at the times shown on the y -axis using synchronized parasites at 1% parasitemia and 1% haematocrit that were incubated for either 27 h (a) or 54 h (b) for P. knowlesi , and 48 h (c) or 96 h (d) for P. falciparum . Signal window (squares) and assay quality ( Z ′ factor, circles) (e–h) were determined as in Figure 1 .

    Journal: Journal of Antimicrobial Chemotherapy

    Article Title: Comparison of the susceptibility of Plasmodium knowlesi and Plasmodium falciparum to antimalarial agents

    doi: 10.1093/jac/dkx279

    Figure Lengend Snippet: Effect of synchrony on drug susceptibility measured across the life cycle using the SYBR Green I method after one or two life cycles. EC 50 values for chloroquine (squares), dihydroartemisinin (circles) or pyrimethamine (diamonds) were determined from experiments initiated at the times shown on the y -axis using synchronized parasites at 1% parasitemia and 1% haematocrit that were incubated for either 27 h (a) or 54 h (b) for P. knowlesi , and 48 h (c) or 96 h (d) for P. falciparum . Signal window (squares) and assay quality ( Z ′ factor, circles) (e–h) were determined as in Figure 1 .

    Article Snippet: Microplates were thawed and incubated with 100 μL of SYBR Green lysis buffer [1:5000 SYBR Green I (Thermo Fisher Scientific, S7563), diluted in 20 mM Tris, 5 mM EDTA, 0.008% (w/v) saponin, 0.08% (v/v) Triton X-100, pH 7.5] in the dark for 1 h, before fluorescence was read in a Spectramax M3 microplate reader (Molecular Devices) at 490 nm excitation and 520 nm emission.

    Techniques: SYBR Green Assay, Incubation

    Influence of starting parasitaemia of P. knowlesi (A1-H.1) and P. falciparum (3D7) on assay quality for both the fluorescent and colorimetric methods. Parasites set to 1% haematocrit and varying parasitaemia (0.1%–2%) were cultured in the presence or absence of a supralethal concentration of chloroquine for 27 h (circles), 54 h (squares) or 81 h (diamonds) for P. knowlesi , and 48 h (cirlces) or 96 h (squares) for P. falciparum . Upon termination of the assay, the plates were read using either the SYBR Green I fluorescence assay (a, c, e and g) or the LDH assay (b, d, f and h). The signal window and Z ′ factor were calculated for each assay. The signal window was calculated by dividing the average reading for the drug-free control by the average reading for the high chloroquine concentration (background) control. The assay quality was assessed by determining the Z ′ factor using the formula described in Zhang et al. 17

    Journal: Journal of Antimicrobial Chemotherapy

    Article Title: Comparison of the susceptibility of Plasmodium knowlesi and Plasmodium falciparum to antimalarial agents

    doi: 10.1093/jac/dkx279

    Figure Lengend Snippet: Influence of starting parasitaemia of P. knowlesi (A1-H.1) and P. falciparum (3D7) on assay quality for both the fluorescent and colorimetric methods. Parasites set to 1% haematocrit and varying parasitaemia (0.1%–2%) were cultured in the presence or absence of a supralethal concentration of chloroquine for 27 h (circles), 54 h (squares) or 81 h (diamonds) for P. knowlesi , and 48 h (cirlces) or 96 h (squares) for P. falciparum . Upon termination of the assay, the plates were read using either the SYBR Green I fluorescence assay (a, c, e and g) or the LDH assay (b, d, f and h). The signal window and Z ′ factor were calculated for each assay. The signal window was calculated by dividing the average reading for the drug-free control by the average reading for the high chloroquine concentration (background) control. The assay quality was assessed by determining the Z ′ factor using the formula described in Zhang et al. 17

    Article Snippet: Microplates were thawed and incubated with 100 μL of SYBR Green lysis buffer [1:5000 SYBR Green I (Thermo Fisher Scientific, S7563), diluted in 20 mM Tris, 5 mM EDTA, 0.008% (w/v) saponin, 0.08% (v/v) Triton X-100, pH 7.5] in the dark for 1 h, before fluorescence was read in a Spectramax M3 microplate reader (Molecular Devices) at 490 nm excitation and 520 nm emission.

    Techniques: Cell Culture, Concentration Assay, SYBR Green Assay, Fluorescence, Lactate Dehydrogenase Assay

    ARRB1 interacts with ASK1 and then antagonizes its TRAF6‐mediated K6‐linked polyubiquitination, thus inhibiting its activation in hepatocytes during hepatic I/R injury. A, Immunoblotting detection of phosphorylated ASK1 protein (left panel) and co‐IP assay for the interaction between His‐ASK1 and HA‐TRAF proteins (right panel) in L02 hepatocytes stimulated with H/R for 6 h. B, His‐tagged ASK1 and HA‐tagged TRAF6 plasmids were cotransfected into L02 hepatocytes. Anti‐His antibody (left panel) and anti‐HA antibody (right panel) were used for immunoprecipitation. C, Immunoprecipitation analysis of indicated ubiquitination types of ASK1 in L02 hepatocytes transfected with HA‐TRAF6 or empty vector in respond to H/R insult or not. D, IP analysis was conducted to detect the binding association between ASK1 and ARRB1 in L02 hepatocytes under normal condition. His‐tagged ASK1 and Flag‐tagged ARRB1 plasmids were cotransfected into L02 hepatocytes. Anti‐His antibody (left panel) and anti‐Flag antibody (right panel) were used for immunoprecipitation. E, IP analysis was conducted to detect the binding association between ASK1 and ARRB1 in L02 hepatocytes when treated with H/R challenge. Anti‐His antibody (left panel) and anti‐Flag antibody (right panel) were used for immunoprecipitation. F, IP analysis showing the expression level of ARRB1 and binding capability between ASK1 and TRAF6 in L02 hepatocytes under H/R challenge or not. Anti‐His antibody (left panel) and anti‐HA antibody (right panel) were used for immunoprecipitation. G and H, Lysates of liver lobes challenged with or without I/R surgery (G) or whole‐cell lysates of primary hepatocytes subjected to or not H/R insult (H) were subjected to immunoprecipitation with anti‐ASK1 antibody followed by immunoblotting with anti‐K6‐linked polyubiquitination antibody when ARRB1 was overexpressed or knockout

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: ARRB1 ameliorates liver ischaemia/reperfusion injury via antagonizing TRAF6‐mediated Lysine 6‐linked polyubiquitination of ASK1 in hepatocytes, et al. ARRB1 ameliorates liver ischaemia/reperfusion injury via antagonizing TRAF6‐mediated Lysine 6‐linked polyubiquitination of ASK1 in hepatocytes

    doi: 10.1111/jcmm.15412

    Figure Lengend Snippet: ARRB1 interacts with ASK1 and then antagonizes its TRAF6‐mediated K6‐linked polyubiquitination, thus inhibiting its activation in hepatocytes during hepatic I/R injury. A, Immunoblotting detection of phosphorylated ASK1 protein (left panel) and co‐IP assay for the interaction between His‐ASK1 and HA‐TRAF proteins (right panel) in L02 hepatocytes stimulated with H/R for 6 h. B, His‐tagged ASK1 and HA‐tagged TRAF6 plasmids were cotransfected into L02 hepatocytes. Anti‐His antibody (left panel) and anti‐HA antibody (right panel) were used for immunoprecipitation. C, Immunoprecipitation analysis of indicated ubiquitination types of ASK1 in L02 hepatocytes transfected with HA‐TRAF6 or empty vector in respond to H/R insult or not. D, IP analysis was conducted to detect the binding association between ASK1 and ARRB1 in L02 hepatocytes under normal condition. His‐tagged ASK1 and Flag‐tagged ARRB1 plasmids were cotransfected into L02 hepatocytes. Anti‐His antibody (left panel) and anti‐Flag antibody (right panel) were used for immunoprecipitation. E, IP analysis was conducted to detect the binding association between ASK1 and ARRB1 in L02 hepatocytes when treated with H/R challenge. Anti‐His antibody (left panel) and anti‐Flag antibody (right panel) were used for immunoprecipitation. F, IP analysis showing the expression level of ARRB1 and binding capability between ASK1 and TRAF6 in L02 hepatocytes under H/R challenge or not. Anti‐His antibody (left panel) and anti‐HA antibody (right panel) were used for immunoprecipitation. G and H, Lysates of liver lobes challenged with or without I/R surgery (G) or whole‐cell lysates of primary hepatocytes subjected to or not H/R insult (H) were subjected to immunoprecipitation with anti‐ASK1 antibody followed by immunoblotting with anti‐K6‐linked polyubiquitination antibody when ARRB1 was overexpressed or knockout

    Article Snippet: Briefly, L02 cells were cotransfected with Flag‐ARRB1 and His‐ASK1 or His‐ASK1 and HA‐TRAF6 for 2 days and sonicated in ice‐cold immunoprecipitation lysis buffer (Thermo Fisher Scientific).

    Techniques: Activation Assay, Co-Immunoprecipitation Assay, Immunoprecipitation, Transfection, Plasmid Preparation, Binding Assay, Expressing, Knock-Out

    Time curve showing mtDNA content measured in a blood sample extracted by BioRobot EZ1 Advanced after incubation using 4 different lysis buffers. The buffers used were G2 (included in the EZ1 Tissue kit) and Pierce RIPA Buffer Lyse (Thermo Fisher Scientific, Waltham, MA, USA). Both buffers were used pure and in combination with Proteinase K (PK). The baseline is obtained using the standard EZ1 tissue kit protocol without incubation.

    Journal: Scientific Reports

    Article Title: Plasmid-normalized quantification of relative mitochondrial DNA copy number

    doi: 10.1038/s41598-018-33684-5

    Figure Lengend Snippet: Time curve showing mtDNA content measured in a blood sample extracted by BioRobot EZ1 Advanced after incubation using 4 different lysis buffers. The buffers used were G2 (included in the EZ1 Tissue kit) and Pierce RIPA Buffer Lyse (Thermo Fisher Scientific, Waltham, MA, USA). Both buffers were used pure and in combination with Proteinase K (PK). The baseline is obtained using the standard EZ1 tissue kit protocol without incubation.

    Article Snippet: After centrifugation (2 min at 2000 × g) the white blood cell pellet obtained was resuspended using two different buffers: the G2 buffer included in the kit and a Pierce RIPA Buffer Lyse (Thermo Fisher Scientific, Waltham, MA, USA).

    Techniques: Incubation, Lysis