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GE Healthcare ecl prime reagents
Ecl Prime Reagents, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 91/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ecl prime reagents/product/GE Healthcare
Average 91 stars, based on 11 article reviews
Price from $9.99 to $1999.99
ecl prime reagents - by Bioz Stars, 2020-07
91/100 stars

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Incubation:

Article Title: Ganglioside GM1 contributes to extracellular/intracellular regulation of insulin resistance, impairment of insulin signaling and down-stream eNOS activation, in human aortic endothelial cells after short- or long-term exposure to TNFα
Article Snippet: .. Membranes were subsequently incubated with the appropriate peroxidase-conjugated secondary antibodies (Cell Signaling Technology), washed and developed with ECL™ Prime reagents (GE Healthcare, Piscataway, NJ, USA). .. The signal was detected using a GE Healthcare ImageQuant LAS3000m.

Article Title: Ganglioside GM2, highly expressed in the MIA PaCa-2 pancreatic ductal adenocarcinoma cell line, is correlated with growth, invasion, and advanced stage
Article Snippet: .. Membranes were then incubated with the appropriate peroxidase-conjugated secondary antibodies (Cell Signaling Technology), washed and developed with ECL™ Prime reagents (GE Healthcare, Piscataway, NJ, USA). .. Heterotopic implantation of PDAC cells In order to clarify in vivo tumorigenicity of GM2– or GM2 + PDAC cells, 1 × 105 cells/animal (n = 5, 5 sites per cell line) were subcutaneously injected into the bilateral flank of nine-week old, female, athymic mice (BALB/cAJcl-nu/nu; CLEA Japan Inc, Tokyo, Japan).

other:

Article Title: Apoptosis-independent cleavage of RhoGDIβ at Asp19 during PMA-stimulated differentiation of THP-1 cells to macrophages
Article Snippet: The immunoreactive proteins were visualized using ECL Prime reagents (GE Healthcare Life Sciences, Ltd., Little Chalfont, UK).

Article Title: Involvement of Macrophages in the Pathogenesis of Familial Amyloid Polyneuropathy and Efficacy of Human iPS Cell-Derived Macrophages in Its Treatment
Article Snippet: The reaction was visualized using ECL prime reagents (GE Healthcare, WI, USA), and detected using a LAS-4000EPUVmini (GE Healthcare).

Article Title: Eros is a novel transmembrane protein that controls the phagocyte respiratory burst and is essential for innate immunity
Article Snippet: Blots were developed using ECL Prime reagents (GE Healthcare) and Lumigen TMA-6.

Article Title: Global Gene Expression Profiling Reveals Functional Importance of Sirt2 in Endothelial Cells under Oxidative Stress
Article Snippet: The blots were developed with ECL Prime reagents from GE Life Sciences (Piscataway, NJ, USA).

Quantitation Assay:

Article Title: Bag1 Co-chaperone Promotes TRC8 E3 Ligase-dependent Degradation of Misfolded Human Ether a Go-Go-related Gene (hERG) Potassium Channels *
Article Snippet: .. Signals were detected using ECL, ECL Plus, or ECL Prime reagents (GE Healthcare), on film, or for quantitation by a FluorChem HD2 digital camera (Alpha Innotech). .. Cells were lysed as above except in PBS containing 0.1% Triton X-100, and lysates were adjusted to the same protein concentration.

Imaging:

Article Title: Body temperature elevation during exercise is essential for activating the Akt signaling pathway in the skeletal muscle of type 2 diabetic rats
Article Snippet: .. After several washes, bands were visualized using ECL Prime reagents (GE Healthcare, Piscataway, NJ, USA) and the signal was recorded with a ChemiDocTM Touch Imaging System (Bio-Rad Laboratories, Hercules, CA, USA). ..

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    GE Healthcare ecl prime western blotting detection reagent
    Semi-quantification of ORF2-3 binding to LMH cells. Different concentrations of ORF2-3 were incubated with cells for 1h. Binding was analyzed by Western blotting applying <t>Amersham™</t> <t>ECL™</t> Prime Western Blotting Detection Reagent.
    Ecl Prime Western Blotting Detection Reagent, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 95/100, based on 1890 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ecl prime western blotting detection reagent/product/GE Healthcare
    Average 95 stars, based on 1890 article reviews
    Price from $9.99 to $1999.99
    ecl prime western blotting detection reagent - by Bioz Stars, 2020-07
    95/100 stars
      Buy from Supplier

    89
    GE Healthcare ecl prime western blotting detection reagent kit
    [ NSI + ] strain contains [ PIN + ] prion which acts as the enhancer of the nonsense suppression. (A) SDD-AGE of protein lysates extracted from the 1-1-D931 [ NSI + ] and 1-1-1-D931 [ nsi - ] strains expressing pCUP1-RNQ1-CFP(LEU2) plasmid. Protein lysates were treated with 1% SDS at room temperature. SDS-resistant aggregates of Rnq1-CFP were detected using monoclonal rabbit primary antibodies against <t>GFP</t> [E385] (ab32146) (Abcam, Great Britain) and <t>ECL</t> Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (B) The effects of RNQ1 deletion on the [ NSI + ] phenotypic manifestation. RNQ1 deletion was obtained as described in Materials and Methods. The [ nsi - ] and [ nsi - ] rnq Δ strains were obtained from the corresponding [ NSI + ] strains by GuHCl treatment. To express RNQ1 , the 5-1-1-D931 [ NSI + ] rnq Δ strain and its [ nsi - ] derivative were transformed with the YGPM25a02 plasmid containing a genomic fragment encoding RNQ1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with an empty vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade medium or–Leu medium with 150 μM CuSO 4 containing galactose as the sole carbon source. Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.
    Ecl Prime Western Blotting Detection Reagent Kit, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 89/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ecl prime western blotting detection reagent kit/product/GE Healthcare
    Average 89 stars, based on 32 article reviews
    Price from $9.99 to $1999.99
    ecl prime western blotting detection reagent kit - by Bioz Stars, 2020-07
    89/100 stars
      Buy from Supplier

    Image Search Results


    Semi-quantification of ORF2-3 binding to LMH cells. Different concentrations of ORF2-3 were incubated with cells for 1h. Binding was analyzed by Western blotting applying Amersham™ ECL™ Prime Western Blotting Detection Reagent.

    Journal: PLoS ONE

    Article Title: C-Terminal Amino Acids 471-507 of Avian Hepatitis E Virus Capsid Protein Are Crucial for Binding to Avian and Human Cells

    doi: 10.1371/journal.pone.0153723

    Figure Lengend Snippet: Semi-quantification of ORF2-3 binding to LMH cells. Different concentrations of ORF2-3 were incubated with cells for 1h. Binding was analyzed by Western blotting applying Amersham™ ECL™ Prime Western Blotting Detection Reagent.

    Article Snippet: Amersham™ ECL™ Prime Western Blotting Detection Reagent (GE Healthcare, Little Chalfont, Bukinghamshire, UK) was applied for semi-quantitative analysis and SuperSignal™ West Pico Chemiluminescent Substrate (Pierce, Thermo Fisher Scientific, Life Technology, Carlsbad, CA, USA) was used for qualitative detection

    Techniques: Binding Assay, Incubation, Western Blot

    Validation of biotin-PK-DPP sensitivity for detection of trypsin-like enzymes ( A) . 1 µM PK-ABP was incubated with an increasing concentration of trypsin and the biotinylated trypsin product was visualized by electrophoresis followed by detection using streptavidin-linked horseradish peroxidase and ECL. ( B ) Trypsin was treated first with the broad-spectrum serine protease inhibitor AEBSF (4 mM) (the + condition) prior to its reaction with ABP and ECL detection.

    Journal: Scientific Reports

    Article Title: Functional Proteomic Profiling of Secreted Serine Proteases in Health and Inflammatory Bowel Disease

    doi: 10.1038/s41598-018-26282-y

    Figure Lengend Snippet: Validation of biotin-PK-DPP sensitivity for detection of trypsin-like enzymes ( A) . 1 µM PK-ABP was incubated with an increasing concentration of trypsin and the biotinylated trypsin product was visualized by electrophoresis followed by detection using streptavidin-linked horseradish peroxidase and ECL. ( B ) Trypsin was treated first with the broad-spectrum serine protease inhibitor AEBSF (4 mM) (the + condition) prior to its reaction with ABP and ECL detection.

    Article Snippet: Membranes were incubated with streptavidin-HRP (Life Technologies), and bands were visualized with ECL Prime Western Blot Detection Reagent (GE Healthcare Life Sciences) and quantified by chemiluminescence yield (Chemidoc XRS; Bio-Rad).

    Techniques: Incubation, Concentration Assay, Electrophoresis, Protease Inhibitor

    Western blot analysis of SjAly from Saccharina japonica . Each sample was applied to one gel in order of symmetry. After electrophoresis, the gel was cut in the middle, one stained with Coomassie Brilliant Blue and the other transferred to a nitrocellulose filter for western blotting. The primary antibody was diluted 1,000 times with 1 × TBS (20 mM Tris-HCl (pH 7.5) and 150 mM NaCl) and used, and the secondary antibody was similarly diluted 10,000 times and used. Protein bands were visualized using an ECL prime western blotting detection reagent (GE Healthcare Life Sciences, Pittsburgh, PA) and chemi-luminescence imager (EZ-capture MG, ATTO, Tokyo, Japan) in an auto exposure mode. An overexposed image is represented in Supplementary Fig. S8 . ( a ) SDS-polyacrylamide gel stained using Coomassie Brilliant Blue. ( b ) Western blot analysis using anti-SjAly antibodies. rSjAly , purified recombinant SjAly; Marker , protein ladder; Blade , protein extract from the blade; Stipe , protein extract from the stipe; Rhizoid , protein extract from the rhizoid. Full-length gel and blot are presented here.

    Journal: Scientific Reports

    Article Title: Functional identification of alginate lyase from the brown alga Saccharina japonica

    doi: 10.1038/s41598-019-41351-6

    Figure Lengend Snippet: Western blot analysis of SjAly from Saccharina japonica . Each sample was applied to one gel in order of symmetry. After electrophoresis, the gel was cut in the middle, one stained with Coomassie Brilliant Blue and the other transferred to a nitrocellulose filter for western blotting. The primary antibody was diluted 1,000 times with 1 × TBS (20 mM Tris-HCl (pH 7.5) and 150 mM NaCl) and used, and the secondary antibody was similarly diluted 10,000 times and used. Protein bands were visualized using an ECL prime western blotting detection reagent (GE Healthcare Life Sciences, Pittsburgh, PA) and chemi-luminescence imager (EZ-capture MG, ATTO, Tokyo, Japan) in an auto exposure mode. An overexposed image is represented in Supplementary Fig. S8 . ( a ) SDS-polyacrylamide gel stained using Coomassie Brilliant Blue. ( b ) Western blot analysis using anti-SjAly antibodies. rSjAly , purified recombinant SjAly; Marker , protein ladder; Blade , protein extract from the blade; Stipe , protein extract from the stipe; Rhizoid , protein extract from the rhizoid. Full-length gel and blot are presented here.

    Article Snippet: The reaction signal was detected using ECL prime western blotting detection reagent (GE Healthcare Life Sciences, Pittsburgh, PA).

    Techniques: Western Blot, Electrophoresis, Staining, Purification, Recombinant, Marker

    [ NSI + ] strain contains [ PIN + ] prion which acts as the enhancer of the nonsense suppression. (A) SDD-AGE of protein lysates extracted from the 1-1-D931 [ NSI + ] and 1-1-1-D931 [ nsi - ] strains expressing pCUP1-RNQ1-CFP(LEU2) plasmid. Protein lysates were treated with 1% SDS at room temperature. SDS-resistant aggregates of Rnq1-CFP were detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (B) The effects of RNQ1 deletion on the [ NSI + ] phenotypic manifestation. RNQ1 deletion was obtained as described in Materials and Methods. The [ nsi - ] and [ nsi - ] rnq Δ strains were obtained from the corresponding [ NSI + ] strains by GuHCl treatment. To express RNQ1 , the 5-1-1-D931 [ NSI + ] rnq Δ strain and its [ nsi - ] derivative were transformed with the YGPM25a02 plasmid containing a genomic fragment encoding RNQ1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with an empty vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade medium or–Leu medium with 150 μM CuSO 4 containing galactose as the sole carbon source. Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.

    Journal: PLoS Genetics

    Article Title: Interaction of Prions Causes Heritable Traits in Saccharomyces cerevisiae

    doi: 10.1371/journal.pgen.1006504

    Figure Lengend Snippet: [ NSI + ] strain contains [ PIN + ] prion which acts as the enhancer of the nonsense suppression. (A) SDD-AGE of protein lysates extracted from the 1-1-D931 [ NSI + ] and 1-1-1-D931 [ nsi - ] strains expressing pCUP1-RNQ1-CFP(LEU2) plasmid. Protein lysates were treated with 1% SDS at room temperature. SDS-resistant aggregates of Rnq1-CFP were detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (B) The effects of RNQ1 deletion on the [ NSI + ] phenotypic manifestation. RNQ1 deletion was obtained as described in Materials and Methods. The [ nsi - ] and [ nsi - ] rnq Δ strains were obtained from the corresponding [ NSI + ] strains by GuHCl treatment. To express RNQ1 , the 5-1-1-D931 [ NSI + ] rnq Δ strain and its [ nsi - ] derivative were transformed with the YGPM25a02 plasmid containing a genomic fragment encoding RNQ1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with an empty vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade medium or–Leu medium with 150 μM CuSO 4 containing galactose as the sole carbon source. Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.

    Article Snippet: Proteins fused with CFP, GFP, and YFP were detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and the Amersham ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA).

    Techniques: Expressing, Plasmid Preparation, Western Blot, Transformation Assay, Incubation

    Mit1 is not a determinant of the [ NSI + ] factor. (A) SDD-AGE assay of protein lysates extracted from the 4-1-1-D931 [ NSI + ] and 1-4-1-1-D931 [ nsi - ] strains expressing pMIT1-MIT1-GFP(URA3) plasmid. Cells were grown for 48 h at 30°C in liquid–Ura selective medium containing 150 μM CuSO 4 . Protein lysates were treated with 1% SDS at room temperature. Mit1-GFP was detected with monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (B) MIT1 deletion does not affect the [ NSI + ] phenotypic manifestation. MIT1 deletion was obtained as described in Materials and Methods. The [ nsi - ] derivative of the 2–936 [ NSI + ] mit1 Δ was obtained by GuHCl treatment. To express MIT1 , 2–936 [ NSI + ] mit1 Δ strain was transformed with YGPM21o12 plasmid from the YSC4613 genomic library containing a genomic fragment encoding MIT1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with a vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade or–Leu Gal media with 150 μM CuSO 4 . Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.

    Journal: PLoS Genetics

    Article Title: Interaction of Prions Causes Heritable Traits in Saccharomyces cerevisiae

    doi: 10.1371/journal.pgen.1006504

    Figure Lengend Snippet: Mit1 is not a determinant of the [ NSI + ] factor. (A) SDD-AGE assay of protein lysates extracted from the 4-1-1-D931 [ NSI + ] and 1-4-1-1-D931 [ nsi - ] strains expressing pMIT1-MIT1-GFP(URA3) plasmid. Cells were grown for 48 h at 30°C in liquid–Ura selective medium containing 150 μM CuSO 4 . Protein lysates were treated with 1% SDS at room temperature. Mit1-GFP was detected with monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (B) MIT1 deletion does not affect the [ NSI + ] phenotypic manifestation. MIT1 deletion was obtained as described in Materials and Methods. The [ nsi - ] derivative of the 2–936 [ NSI + ] mit1 Δ was obtained by GuHCl treatment. To express MIT1 , 2–936 [ NSI + ] mit1 Δ strain was transformed with YGPM21o12 plasmid from the YSC4613 genomic library containing a genomic fragment encoding MIT1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with a vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade or–Leu Gal media with 150 μM CuSO 4 . Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.

    Article Snippet: Proteins fused with CFP, GFP, and YFP were detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and the Amersham ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA).

    Techniques: Expressing, Plasmid Preparation, Western Blot, Transformation Assay, Incubation

    [ SWI + ] prion is a key determinant of nonsense suppression in [ NSI + ] strains. (A) Sedimentation analysis of Swi1(1–297)-YFP protein from the 4-1-1-D931 [ NSI + ] and 1-4-1-1-D931 [ nsi - ] strains expressing pCUP1-SWI1(1–297)-YFP (URA3) plasmid. Soluble (S) and insoluble (I) fractions were obtained as indicated in Materials and Methods. Swi1(1–297)-YFP was detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). Next, SDD-AGE analysis of insoluble fractions of [ NSI + ] and [ nsi - ] strains comprising Swi1(1–297)-YFP was performed. (B) The effects of SWI1 deletion on the [ NSI + ] phenotypic manifestation. SWI1 deletion was obtained as described in Materials and Methods. To express SWI1 , the 11-1-1-D931 [ NSI + ] swi1 Δ strain was transformed with the YGPM19p21 plasmid from the YSC4613 genomic library, containing a genomic fragment encoding SWI1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with an empty vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade or–Leu Gal media with 150 μM CuSO 4 . Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.

    Journal: PLoS Genetics

    Article Title: Interaction of Prions Causes Heritable Traits in Saccharomyces cerevisiae

    doi: 10.1371/journal.pgen.1006504

    Figure Lengend Snippet: [ SWI + ] prion is a key determinant of nonsense suppression in [ NSI + ] strains. (A) Sedimentation analysis of Swi1(1–297)-YFP protein from the 4-1-1-D931 [ NSI + ] and 1-4-1-1-D931 [ nsi - ] strains expressing pCUP1-SWI1(1–297)-YFP (URA3) plasmid. Soluble (S) and insoluble (I) fractions were obtained as indicated in Materials and Methods. Swi1(1–297)-YFP was detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). Next, SDD-AGE analysis of insoluble fractions of [ NSI + ] and [ nsi - ] strains comprising Swi1(1–297)-YFP was performed. (B) The effects of SWI1 deletion on the [ NSI + ] phenotypic manifestation. SWI1 deletion was obtained as described in Materials and Methods. To express SWI1 , the 11-1-1-D931 [ NSI + ] swi1 Δ strain was transformed with the YGPM19p21 plasmid from the YSC4613 genomic library, containing a genomic fragment encoding SWI1 under the control of its endogenous promoter. Other strains presented in this Figure were transformed with an empty vector expressing only the LEU2 gene. Transformants were selected on–Leu medium with 150 μM CuSO 4 and replica-plated on–Leu–Ade or–Leu Gal media with 150 μM CuSO 4 . Images were taken after 5 days of incubation of–Ade plates or after 3 passages on Gal plates.

    Article Snippet: Proteins fused with CFP, GFP, and YFP were detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and the Amersham ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA).

    Techniques: Sedimentation, Expressing, Plasmid Preparation, Western Blot, Transformation Assay, Incubation

    [ SWI + ] and [ PIN + ] prions demonstrate complementary interaction. (A) Comparative analysis of the growth of strains containing combinations of [ prion - ] or [ PRION + ] states for Rnq1 and Swi1 as well as deletions of the corresponding genes. “1”–The [ SWI + ][ pin - ] and [ swi - ][ PIN + ] strains were obtained from the 1-1-D931 [ SWI + ][ PIN + ] strain by deletion with subsequent reintroduction of RNQ1 and SWI1 genes, respectively (see “ Materials and Methods ”). The [ swi - ][ pin - ] strain was obtained from the 1-1-D931 [ SWI + ][ PIN + ] strain by GuHCl curing. “2”–The 26-1-4-1-1-D931 [ swi - ][ PIN + ], 12-1-4-1-1-D931 [ SWI + ][ pin - ], and 16-1-4-1-1-D931 [ SWI + ][ PIN + ] strains were obtained by transformation of the 1-4-1-1-D931 [ swi - ][ pin - ] recipient yeast cells with the 1-1-D931 [ SWI + ][ PIN + ] protein lysates followed by analysis of [ SWI ] and [ PIN ] status of the cells as described in “Materials and Methods”. Images were obtained after 5 days of incubation on–Ade plates with 150 μM CuSO 4 or after 3 passages on Gal plates. (B) Sedimentation analysis of Swi1(1–297)-YFP protein from the 16-1-4-1-1-D931 [ SWI + ][ PIN + ] and 12-1-4-1-1-D931 [ SWI + ][ pin - ] strains expressing the pCUP1-SWI1(1–297)-YFP (URA3) plasmid. Soluble (S) and insoluble (I) fractions were obtained as indicated in Materials and Methods. Swi1(1–297)-YFP was detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (C) Analysis of the colocalization of Swi1-YFP and Rnq1-CFP aggregates. The cells of the D938 [ SWI + ][ PIN + ] strain were co-transformed with p426GPD–SWI1YFP and pCUP1-RNQ1-CFP(LEU2) plasmids. Transformants were selected on–Ura–Leu selective media with 150 μM CuSO 4 and incubated for 48 h prior to fluorescence microscopy.

    Journal: PLoS Genetics

    Article Title: Interaction of Prions Causes Heritable Traits in Saccharomyces cerevisiae

    doi: 10.1371/journal.pgen.1006504

    Figure Lengend Snippet: [ SWI + ] and [ PIN + ] prions demonstrate complementary interaction. (A) Comparative analysis of the growth of strains containing combinations of [ prion - ] or [ PRION + ] states for Rnq1 and Swi1 as well as deletions of the corresponding genes. “1”–The [ SWI + ][ pin - ] and [ swi - ][ PIN + ] strains were obtained from the 1-1-D931 [ SWI + ][ PIN + ] strain by deletion with subsequent reintroduction of RNQ1 and SWI1 genes, respectively (see “ Materials and Methods ”). The [ swi - ][ pin - ] strain was obtained from the 1-1-D931 [ SWI + ][ PIN + ] strain by GuHCl curing. “2”–The 26-1-4-1-1-D931 [ swi - ][ PIN + ], 12-1-4-1-1-D931 [ SWI + ][ pin - ], and 16-1-4-1-1-D931 [ SWI + ][ PIN + ] strains were obtained by transformation of the 1-4-1-1-D931 [ swi - ][ pin - ] recipient yeast cells with the 1-1-D931 [ SWI + ][ PIN + ] protein lysates followed by analysis of [ SWI ] and [ PIN ] status of the cells as described in “Materials and Methods”. Images were obtained after 5 days of incubation on–Ade plates with 150 μM CuSO 4 or after 3 passages on Gal plates. (B) Sedimentation analysis of Swi1(1–297)-YFP protein from the 16-1-4-1-1-D931 [ SWI + ][ PIN + ] and 12-1-4-1-1-D931 [ SWI + ][ pin - ] strains expressing the pCUP1-SWI1(1–297)-YFP (URA3) plasmid. Soluble (S) and insoluble (I) fractions were obtained as indicated in Materials and Methods. Swi1(1–297)-YFP was detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA). (C) Analysis of the colocalization of Swi1-YFP and Rnq1-CFP aggregates. The cells of the D938 [ SWI + ][ PIN + ] strain were co-transformed with p426GPD–SWI1YFP and pCUP1-RNQ1-CFP(LEU2) plasmids. Transformants were selected on–Ura–Leu selective media with 150 μM CuSO 4 and incubated for 48 h prior to fluorescence microscopy.

    Article Snippet: Proteins fused with CFP, GFP, and YFP were detected using monoclonal rabbit primary antibodies against GFP [E385] (ab32146) (Abcam, Great Britain) and the Amersham ECL Prime Western Blotting Detection Reagent kit (GE Healthcare, USA).

    Techniques: Transformation Assay, Incubation, Sedimentation, Expressing, Plasmid Preparation, Western Blot, Fluorescence, Microscopy