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GE Healthcare ecl prime
Ecl Prime, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 99/100, based on 431 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/ecl prime/product/GE Healthcare
Average 99 stars, based on 431 article reviews
Price from $9.99 to $1999.99
ecl prime - by Bioz Stars, 2020-02
99/100 stars

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Related Articles

Blocking Assay:

Article Title: Annexin A5 is the Most Abundant Membrane-Associated Protein in Stereocilia but is Dispensable for Hair-Bundle Development and Function
Article Snippet: .. Membranes were blocked with Prime Blocking Agent (GE Healthcare Life Sciences RPN418), and probed with specific primary antibodies which were detected with species-specific HRP-coupled secondary antibodies (Jackson ImmunoResearch AffiniPure 111-035-144, and 115-035-003) and ECL Prime (GE Healthcare Life Sciences RPN2232). .. Vestibular evoked potentials For VsEP testing, mice were anesthetized with a solution containing ketamine (18 mg/ml) and xylazine (2 mg/ml), using 5–9 μl per gram body weight injected intraperitoneally.

Article Title: Dengue viruses cleave STING in humans but not in nonhuman primates, their presumed natural reservoir
Article Snippet: Blocking was performed with a 10% milk solution in tris-buffered saline supplemented with 0.1% TWEEN20. .. Blots were developed using ECL Prime (Amersham RPN2232) and imaged using ImagQuant LAS 4000 (Amersham 28-9558-10).

Article Title: BubR1 Insufficiency Results in Decreased Macrophage Proliferation and Attenuated Atherogenesis in Apolipoprotein E‐Deficient Mice
Article Snippet: The proteins were transferred onto nitrocellulose membranes, and probed with the specified primary antibodies diluted in 4% (w/v) Block Ace (UKB80; DS Pharma Biomedical Co., Ltd, Osaka, Japan) in TBS 0.1% Tween20. .. Antibody‐labeled proteins were visualized using enhanced chemiluminescence, ECL prime (RPN2232; GE Healthcare).

Electrophoresis:

Article Title: Cancer reversion with oocyte extracts is mediated by cell cycle arrest and induction of tumour dormancy
Article Snippet: Proteins were loaded into 8-12% polyacrylamide gels (30 μg/lane), separated by SDS-PAGE electrophoresis and blotted onto a PVDF membrane. .. ECL Prime (GE Healthcare) was used to detect chemiluminescence.

Incubation:

Article Title: Cancer reversion with oocyte extracts is mediated by cell cycle arrest and induction of tumour dormancy
Article Snippet: Peroxidase conjugated secondary antibodies were incubated for 1h at RT. .. ECL Prime (GE Healthcare) was used to detect chemiluminescence.

Article Title: Krüppel-like factor 6 is a transcriptional activator of autophagy in acute liver injury
Article Snippet: .. After incubation with the appropriate horseradish peroxidase-conjugated secondary antibody, bound antibodies were visualized using ECL-Prime (GE Healthcare, Chalfont St. Giles, UK). .. Blotting images were generated using ChemiDoc System and Quantity One software (Bio-Rad) to quantify the densities of the bands.

Article Title: Pathological significance and prognostic significance of FES expression in bladder cancer vary according to tumor grade
Article Snippet: After three washes with TBST, membranes were incubated with the appropriate secondary antibody for 1 h at room temperature. .. Specific protein bands were visualized using ECL Prime (GE Healthcare, Little Chalfont, UK).

Article Title: A huntingtin knock-in pig model recapitulates features of selective neurodegeneration in Huntington’s disease
Article Snippet: The blotted membran e was washed in TBST 3 times (each time 5 min), followed by incubation with HRP-conjugated secondary antibodies in 5% milk/TBST for 1 h at room temperature. .. After three washes in TBST, ECL Prime (GE Healthcare) was used to detect immunoreactive signals on the membrane.

Article Title: Modulation of Human Leukocyte Antigen-C by Human Cytomegalovirus Stimulates KIR2DS1 Recognition by Natural Killer Cells
Article Snippet: The membranes were incubated overnight at 4°C or 3 h at room temperature with primary antibodies [anti-pp28 (Abcam), L31, HC10, anti-I.E.1 (Merck Millipore, Billerica, MA, USA), anti-Calnexin (Enzo Life Sciences, Farmingdale, NY, USA), and anti-Flag (M2, Sigma-Aldrich)] at concentration of 0.1–0.2 μg. .. Chemiluminescence was performed according to the manufacturer’s instructions using ECL Prime (GE Healthcare) or home-made ECL ( ).

Expressing:

Article Title: Prolyl-hydroxylase inhibition induces SDF-1 associated with increased CXCR4+/CD11b+ subpopulations and cardiac repair
Article Snippet: Appropriate horseradish-peroxidase-coupled secondary antibodies were used, and chemiluminescence was performed with ECL-Prime (Amersham). .. HIF-1α protein expression was normalized to GAPDH (cytosol) and TBP (nuclear compartment).

Bradford Assay:

Article Title: Death-associated protein kinase 1 has a critical role in aberrant tau protein regulation and function
Article Snippet: The soluble protein concentration was determined by Bradford assay (Bio-Rad). .. Antibody-bound proteins were detected using the Western Lightning Plus-ECL (PerkinElmer) or ECL Prime (GE Healthcare Life Sciences, Pittsburgh, PA, USA) chemiluminescence system.

Western Blot:

Article Title: RACK-1 regulates let-7 microRNA expression and terminal cell differentiation in Caenorhabditis elegans
Article Snippet: Paragraph title: Antibodies and western blotting ... Horseradish peroxidase-conjugated anti-rabbit (211-032-171, Jackson ImmunoResearch Labs, 1:10 000 dilution) or anti-mouse (115-035-174, Jackson ImmunoResearch Labs, 1:10 000 dilution) light-chain-specific secondary antibodies were used for signal detection by ECL Prime (GE Healthcare).

Article Title: Cancer reversion with oocyte extracts is mediated by cell cycle arrest and induction of tumour dormancy
Article Snippet: Paragraph title: Western blotting ... ECL Prime (GE Healthcare) was used to detect chemiluminescence.

Article Title: Prolyl-hydroxylase inhibition induces SDF-1 associated with increased CXCR4+/CD11b+ subpopulations and cardiac repair
Article Snippet: Paragraph title: Western blot ... Appropriate horseradish-peroxidase-coupled secondary antibodies were used, and chemiluminescence was performed with ECL-Prime (Amersham).

Article Title: Krüppel-like factor 6 is a transcriptional activator of autophagy in acute liver injury
Article Snippet: Paragraph title: Western blotting ... After incubation with the appropriate horseradish peroxidase-conjugated secondary antibody, bound antibodies were visualized using ECL-Prime (GE Healthcare, Chalfont St. Giles, UK).

Article Title: H19-Dependent Transcriptional Regulation of β3 and β4 Integrins Upon Estrogen and Hypoxia Favors Metastatic Potential in Prostate Cancer
Article Snippet: Paragraph title: 4.8. Protein Extraction and Western Blot ... Specific protein signals were revealed with ECL Prime (Amersham, GE Healthcare, Boston, MA, USA) and detected by UVIDOC (Eppendorf S.r.l., Hamburg, Germany).

Article Title: Pathological significance and prognostic significance of FES expression in bladder cancer vary according to tumor grade
Article Snippet: Paragraph title: Western blot analysis ... Specific protein bands were visualized using ECL Prime (GE Healthcare, Little Chalfont, UK).

Article Title: Dengue viruses cleave STING in humans but not in nonhuman primates, their presumed natural reservoir
Article Snippet: Paragraph title: Western blotting ... Blots were developed using ECL Prime (Amersham RPN2232) and imaged using ImagQuant LAS 4000 (Amersham 28-9558-10).

Article Title: A huntingtin knock-in pig model recapitulates features of selective neurodegeneration in Huntington’s disease
Article Snippet: Paragraph title: Western blot analysis, immunohistochemistry, and electron microscopy ... After three washes in TBST, ECL Prime (GE Healthcare) was used to detect immunoreactive signals on the membrane.

Article Title: Death-associated protein kinase 1 has a critical role in aberrant tau protein regulation and function
Article Snippet: .. Antibody-bound proteins were detected using the Western Lightning Plus-ECL (PerkinElmer) or ECL Prime (GE Healthcare Life Sciences, Pittsburgh, PA, USA) chemiluminescence system. .. The following antibodies were used to probe different proteins: Flag, DAPK1 (Sigma); DCX, β -actin, α -tubulin (Cell Signaling Technology, Danvers, MA, USA); PHF-13 (p-Ser396-Tau) (Anaspec, Fremont, CA, USA); Tau-5, p-Tau (Ser262); and AT180 (pThr231-Tau) (Invitrogen).

Article Title: Transcriptome profiling reveals novel BMI- and sex-specific gene expression signatures for human cardiac hypertrophy
Article Snippet: For Western blotting procedures, protein concentrations were normalized between samples to 10–30 μg and mixed with Non-Reducing Lane Marker (Thermo Scientific) with 5% β-mercaptoethanol. .. After five washes with TBS-T, blots were developed with a standard ECL kit (Life Technologies) or ECL Prime (Amersham) on X-ray film or using a G:BOX digital imaging system (Syngene).

Article Title: BubR1 Insufficiency Results in Decreased Macrophage Proliferation and Attenuated Atherogenesis in Apolipoprotein E‐Deficient Mice
Article Snippet: Paragraph title: Western Blotting ... Antibody‐labeled proteins were visualized using enhanced chemiluminescence, ECL prime (RPN2232; GE Healthcare).

Article Title: p120RasGAP Is a Mediator of Rho Pathway Activation and Tumorigenicity in the DLD1 Colorectal Cancer Cell Line
Article Snippet: Paragraph title: Immunoprecipitation and Western Blotting ... Westerns were probed with appropriate secondary antibody, reacted with ECL prime (GE Healthcare, Piscataway, NJ) and exposed to XRAY film for the appropriate amount of time.

Article Title: CSB interacts with SNM1A and promotes DNA interstrand crosslink processing
Article Snippet: Paragraph title: Western blotting ... Secondary horseradish peroxidase-conjugated antibodies and ECL prime (GE Healthcare Bio-Sciences) or SuperSignal West Femto Chemiluminescent Substrate (Thermo Fisher Scientific) were used to visualize proteins on a ChemiDoc XRS+ system (Bio-Rad Laboratories, Hercules, CA, USA).

Article Title: Modulation of Human Leukocyte Antigen-C by Human Cytomegalovirus Stimulates KIR2DS1 Recognition by Natural Killer Cells
Article Snippet: Paragraph title: Western Blot ... Chemiluminescence was performed according to the manufacturer’s instructions using ECL Prime (GE Healthcare) or home-made ECL ( ).

Electron Microscopy:

Article Title: A huntingtin knock-in pig model recapitulates features of selective neurodegeneration in Huntington’s disease
Article Snippet: Paragraph title: Western blot analysis, immunohistochemistry, and electron microscopy ... After three washes in TBST, ECL Prime (GE Healthcare) was used to detect immunoreactive signals on the membrane.

Immunohistochemistry:

Article Title: A huntingtin knock-in pig model recapitulates features of selective neurodegeneration in Huntington’s disease
Article Snippet: Paragraph title: Western blot analysis, immunohistochemistry, and electron microscopy ... After three washes in TBST, ECL Prime (GE Healthcare) was used to detect immunoreactive signals on the membrane.

Immunoprecipitation:

Article Title: p120RasGAP Is a Mediator of Rho Pathway Activation and Tumorigenicity in the DLD1 Colorectal Cancer Cell Line
Article Snippet: Paragraph title: Immunoprecipitation and Western Blotting ... Westerns were probed with appropriate secondary antibody, reacted with ECL prime (GE Healthcare, Piscataway, NJ) and exposed to XRAY film for the appropriate amount of time.

Protease Inhibitor:

Article Title: A huntingtin knock-in pig model recapitulates features of selective neurodegeneration in Huntington’s disease
Article Snippet: For western blotting, pig brain tissues were lysed in ice-cold RIPA buffer (50 mM Tris, pH 8.0, 150 mM NaCl, 1 mM EDTA pH 8.0, 1 mM EGTA pH 8.0, 0.1% SDS, 0.5% DOC, and 1% Triton X-100) containing Halt Protease Inhibitor cocktail (Thermo Scientific) and PMSF. .. After three washes in TBST, ECL Prime (GE Healthcare) was used to detect immunoreactive signals on the membrane.

Article Title: Death-associated protein kinase 1 has a critical role in aberrant tau protein regulation and function
Article Snippet: Mouse and human brain extracts were prepared by homogenizing tissue in ice-cold lysis buffer (10 mM Tris-HCl (pH 7.4), 0.8 M NaCl, 1 mM EGTA, 10% sucrose, 1 mM DTT) supplemented with protease inhibitor cocktail and phosphatase inhibitor cocktail. .. Antibody-bound proteins were detected using the Western Lightning Plus-ECL (PerkinElmer) or ECL Prime (GE Healthcare Life Sciences, Pittsburgh, PA, USA) chemiluminescence system.

Qubit Protein Assay:

Article Title: Cancer reversion with oocyte extracts is mediated by cell cycle arrest and induction of tumour dormancy
Article Snippet: The protein content was quantified using a Qubit Protein Assay Kit (Life Technologies) following the manufacturer’s protocol. .. ECL Prime (GE Healthcare) was used to detect chemiluminescence.

Cell Culture:

Article Title: Pathological significance and prognostic significance of FES expression in bladder cancer vary according to tumor grade
Article Snippet: Briefly, cultured cells were harvested by scraping and were lysed in ice-cold hypotonic cell lysis buffer containing protease inhibitors. .. Specific protein bands were visualized using ECL Prime (GE Healthcare, Little Chalfont, UK).

Generated:

Article Title: Krüppel-like factor 6 is a transcriptional activator of autophagy in acute liver injury
Article Snippet: After incubation with the appropriate horseradish peroxidase-conjugated secondary antibody, bound antibodies were visualized using ECL-Prime (GE Healthcare, Chalfont St. Giles, UK). .. Blotting images were generated using ChemiDoc System and Quantity One software (Bio-Rad) to quantify the densities of the bands.

Imaging:

Article Title: RACK-1 regulates let-7 microRNA expression and terminal cell differentiation in Caenorhabditis elegans
Article Snippet: Horseradish peroxidase-conjugated anti-rabbit (211-032-171, Jackson ImmunoResearch Labs, 1:10 000 dilution) or anti-mouse (115-035-174, Jackson ImmunoResearch Labs, 1:10 000 dilution) light-chain-specific secondary antibodies were used for signal detection by ECL Prime (GE Healthcare). .. Images were acquired by UVP BioSpectrum 500 imaging system.

Article Title: Transcriptome profiling reveals novel BMI- and sex-specific gene expression signatures for human cardiac hypertrophy
Article Snippet: .. After five washes with TBS-T, blots were developed with a standard ECL kit (Life Technologies) or ECL Prime (Amersham) on X-ray film or using a G:BOX digital imaging system (Syngene). .. For Western blots, data are shown as means ± SE; Student’s t -test was used for statistical analysis with P < 0.05 being considered as statistically significant, marked with the symbol *.

Protein Concentration:

Article Title: Death-associated protein kinase 1 has a critical role in aberrant tau protein regulation and function
Article Snippet: The soluble protein concentration was determined by Bradford assay (Bio-Rad). .. Antibody-bound proteins were detected using the Western Lightning Plus-ECL (PerkinElmer) or ECL Prime (GE Healthcare Life Sciences, Pittsburgh, PA, USA) chemiluminescence system.

Article Title: Transcriptome profiling reveals novel BMI- and sex-specific gene expression signatures for human cardiac hypertrophy
Article Snippet: Supernatants were placed into new tubes, and protein concentration was recorded with Bradford’s method on an Eppendorf Biophotometer. .. After five washes with TBS-T, blots were developed with a standard ECL kit (Life Technologies) or ECL Prime (Amersham) on X-ray film or using a G:BOX digital imaging system (Syngene).

Article Title: p120RasGAP Is a Mediator of Rho Pathway Activation and Tumorigenicity in the DLD1 Colorectal Cancer Cell Line
Article Snippet: Whole cell lyates were normalized for protein concentration, combined with 6xSDS sample buffer and boiled for 5 minutes as well. .. Westerns were probed with appropriate secondary antibody, reacted with ECL prime (GE Healthcare, Piscataway, NJ) and exposed to XRAY film for the appropriate amount of time.

Binding Assay:

Article Title: Prolyl-hydroxylase inhibition induces SDF-1 associated with increased CXCR4+/CD11b+ subpopulations and cardiac repair
Article Snippet: SDS-PAGE and blotting analysis were done using the following antibodies: HIF-1α (rabbit polyclonal against HIF-1α, ab82832 Abcam), TATA binding protein (TBP) (mouse monoclonal against TBP, ab51841, Abcam), and GAPDH (mouse monoclonal to GAPDH, ab8245, Abcam). .. Appropriate horseradish-peroxidase-coupled secondary antibodies were used, and chemiluminescence was performed with ECL-Prime (Amersham).

Isolation:

Article Title: A huntingtin knock-in pig model recapitulates features of selective neurodegeneration in Huntington’s disease
Article Snippet: After three washes in TBST, ECL Prime (GE Healthcare) was used to detect immunoreactive signals on the membrane. .. For immunohistochemistry, isolated pig brain tissues were fixed 24 hours in 4% paraformaldehyde in 0.01 M PBS and then transferred into 30% sucrose to dehydrate at 4°C.

Size-exclusion Chromatography:

Article Title: A huntingtin knock-in pig model recapitulates features of selective neurodegeneration in Huntington’s disease
Article Snippet: After three washes in TBST, ECL Prime (GE Healthcare) was used to detect immunoreactive signals on the membrane. .. The pig brain tissues were sec tioned at 20 μm using a freezing microtome.

Purification:

Article Title: Dengue viruses cleave STING in humans but not in nonhuman primates, their presumed natural reservoir
Article Snippet: Primary antibodies used were used against HA (3f10 clone Sigma 11867423001), Flag (M2 clone Sigma F3165), GAPDH (CellSignaling 14C10), STING (Abcam 92605), actin (Santa Cruz Biotech Sc47778), and dengue virus NS3 (mouse polyclonal antibody raised against purified full-length NS3 from dengue 2 strain 16681 [ ]). .. Blots were developed using ECL Prime (Amersham RPN2232) and imaged using ImagQuant LAS 4000 (Amersham 28-9558-10).

Protein Extraction:

Article Title: H19-Dependent Transcriptional Regulation of β3 and β4 Integrins Upon Estrogen and Hypoxia Favors Metastatic Potential in Prostate Cancer
Article Snippet: Paragraph title: 4.8. Protein Extraction and Western Blot ... Specific protein signals were revealed with ECL Prime (Amersham, GE Healthcare, Boston, MA, USA) and detected by UVIDOC (Eppendorf S.r.l., Hamburg, Germany).

SDS Page:

Article Title: Cancer reversion with oocyte extracts is mediated by cell cycle arrest and induction of tumour dormancy
Article Snippet: Proteins were loaded into 8-12% polyacrylamide gels (30 μg/lane), separated by SDS-PAGE electrophoresis and blotted onto a PVDF membrane. .. ECL Prime (GE Healthcare) was used to detect chemiluminescence.

Article Title: Annexin A5 is the Most Abundant Membrane-Associated Protein in Stereocilia but is Dispensable for Hair-Bundle Development and Function
Article Snippet: Samples were heated at 95 °C for 5 min, and resolved using 4–12% SDS-PAGE gels with MES or MOPS buffer (NuPAGE gels and buffers, Life Technologies). .. Membranes were blocked with Prime Blocking Agent (GE Healthcare Life Sciences RPN418), and probed with specific primary antibodies which were detected with species-specific HRP-coupled secondary antibodies (Jackson ImmunoResearch AffiniPure 111-035-144, and 115-035-003) and ECL Prime (GE Healthcare Life Sciences RPN2232).

Article Title: Prolyl-hydroxylase inhibition induces SDF-1 associated with increased CXCR4+/CD11b+ subpopulations and cardiac repair
Article Snippet: SDS-PAGE and blotting analysis were done using the following antibodies: HIF-1α (rabbit polyclonal against HIF-1α, ab82832 Abcam), TATA binding protein (TBP) (mouse monoclonal against TBP, ab51841, Abcam), and GAPDH (mouse monoclonal to GAPDH, ab8245, Abcam). .. Appropriate horseradish-peroxidase-coupled secondary antibodies were used, and chemiluminescence was performed with ECL-Prime (Amersham).

Article Title: Krüppel-like factor 6 is a transcriptional activator of autophagy in acute liver injury
Article Snippet: Western blotting For SDS-PAGE 30 µg of total protein were separated; immunoblotting was performed using standard procedures with the following primary antibodies: LC3 (Abcam), KLF6-R173 (Santa Cruz Biotechnologies), p62 (Enzo Lifesciences, Antwerpen, Belgium), β-Actin 13E5 and GAPDH 14C10 (Cell Signaling). .. After incubation with the appropriate horseradish peroxidase-conjugated secondary antibody, bound antibodies were visualized using ECL-Prime (GE Healthcare, Chalfont St. Giles, UK).

Article Title: A huntingtin knock-in pig model recapitulates features of selective neurodegeneration in Huntington’s disease
Article Snippet: The supernatants were loaded to SDS-PAGE, transferred to a PVDF membrane, and blocked with 5% milk/PBS for 1 h at room temperature. .. After three washes in TBST, ECL Prime (GE Healthcare) was used to detect immunoreactive signals on the membrane.

Article Title: Death-associated protein kinase 1 has a critical role in aberrant tau protein regulation and function
Article Snippet: Protein samples (5–10 μ g) were separated by SDS-PAGE and transferred to polyvinylidene fluoride membranes (PerkinElmer, Waltham, MA, USA). .. Antibody-bound proteins were detected using the Western Lightning Plus-ECL (PerkinElmer) or ECL Prime (GE Healthcare Life Sciences, Pittsburgh, PA, USA) chemiluminescence system.

Article Title: BubR1 Insufficiency Results in Decreased Macrophage Proliferation and Attenuated Atherogenesis in Apolipoprotein E‐Deficient Mice
Article Snippet: Western Blotting Cell lysates were dissolved by boiling in Laemmli sample buffer (C3228; Sigma‐Aldrich Ireland Limited, Arklow, Ireland) and proteins were separated using SDS‐PAGE on 7.5% (w/v) polyacrylamide gels. .. Antibody‐labeled proteins were visualized using enhanced chemiluminescence, ECL prime (RPN2232; GE Healthcare).

Article Title: Modulation of Human Leukocyte Antigen-C by Human Cytomegalovirus Stimulates KIR2DS1 Recognition by Natural Killer Cells
Article Snippet: Total cell or virus particle lysates were loaded onto a 10–13% SDS-PAGE gel, and proteins were transferred onto Immobilon-P PVDF membranes (Millipore). .. Chemiluminescence was performed according to the manufacturer’s instructions using ECL Prime (GE Healthcare) or home-made ECL ( ).

Software:

Article Title: Krüppel-like factor 6 is a transcriptional activator of autophagy in acute liver injury
Article Snippet: After incubation with the appropriate horseradish peroxidase-conjugated secondary antibody, bound antibodies were visualized using ECL-Prime (GE Healthcare, Chalfont St. Giles, UK). .. Blotting images were generated using ChemiDoc System and Quantity One software (Bio-Rad) to quantify the densities of the bands.

Article Title: H19-Dependent Transcriptional Regulation of β3 and β4 Integrins Upon Estrogen and Hypoxia Favors Metastatic Potential in Prostate Cancer
Article Snippet: Specific protein signals were revealed with ECL Prime (Amersham, GE Healthcare, Boston, MA, USA) and detected by UVIDOC (Eppendorf S.r.l., Hamburg, Germany). .. The intensity of each band was evaluated using the NIH Image J 1.8 software (National Institutes of Health, Bethesda, MD, USA).

Article Title: CSB interacts with SNM1A and promotes DNA interstrand crosslink processing
Article Snippet: Secondary horseradish peroxidase-conjugated antibodies and ECL prime (GE Healthcare Bio-Sciences) or SuperSignal West Femto Chemiluminescent Substrate (Thermo Fisher Scientific) were used to visualize proteins on a ChemiDoc XRS+ system (Bio-Rad Laboratories, Hercules, CA, USA). .. Digitized images were obtained and quantified with ImageLab version 5.1 (Bio-Rad Laboratories), and processed using the Adobe Photoshop 8.0 software package (Adobe Systems, San Jose, CA, USA).

Concentration Assay:

Article Title: Modulation of Human Leukocyte Antigen-C by Human Cytomegalovirus Stimulates KIR2DS1 Recognition by Natural Killer Cells
Article Snippet: The membranes were incubated overnight at 4°C or 3 h at room temperature with primary antibodies [anti-pp28 (Abcam), L31, HC10, anti-I.E.1 (Merck Millipore, Billerica, MA, USA), anti-Calnexin (Enzo Life Sciences, Farmingdale, NY, USA), and anti-Flag (M2, Sigma-Aldrich)] at concentration of 0.1–0.2 μg. .. Chemiluminescence was performed according to the manufacturer’s instructions using ECL Prime (GE Healthcare) or home-made ECL ( ).

Marker:

Article Title: Transcriptome profiling reveals novel BMI- and sex-specific gene expression signatures for human cardiac hypertrophy
Article Snippet: For Western blotting procedures, protein concentrations were normalized between samples to 10–30 μg and mixed with Non-Reducing Lane Marker (Thermo Scientific) with 5% β-mercaptoethanol. .. After five washes with TBS-T, blots were developed with a standard ECL kit (Life Technologies) or ECL Prime (Amersham) on X-ray film or using a G:BOX digital imaging system (Syngene).

Lysis:

Article Title: Pathological significance and prognostic significance of FES expression in bladder cancer vary according to tumor grade
Article Snippet: Briefly, cultured cells were harvested by scraping and were lysed in ice-cold hypotonic cell lysis buffer containing protease inhibitors. .. Specific protein bands were visualized using ECL Prime (GE Healthcare, Little Chalfont, UK).

Article Title: Death-associated protein kinase 1 has a critical role in aberrant tau protein regulation and function
Article Snippet: Mouse and human brain extracts were prepared by homogenizing tissue in ice-cold lysis buffer (10 mM Tris-HCl (pH 7.4), 0.8 M NaCl, 1 mM EGTA, 10% sucrose, 1 mM DTT) supplemented with protease inhibitor cocktail and phosphatase inhibitor cocktail. .. Antibody-bound proteins were detected using the Western Lightning Plus-ECL (PerkinElmer) or ECL Prime (GE Healthcare Life Sciences, Pittsburgh, PA, USA) chemiluminescence system.

Article Title: p120RasGAP Is a Mediator of Rho Pathway Activation and Tumorigenicity in the DLD1 Colorectal Cancer Cell Line
Article Snippet: Immunoprecipitated proteins were then washed 3x with lysis buffer, eluted with 20 µL of 2xSDS sample buffer and boiled for 5 minutes. .. Westerns were probed with appropriate secondary antibody, reacted with ECL prime (GE Healthcare, Piscataway, NJ) and exposed to XRAY film for the appropriate amount of time.

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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: 99/100, based on 1020 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 99 stars, based on 1020 article reviews
    Price from $9.99 to $1999.99
    ecl prime western blotting detection reagent - by Bioz Stars, 2020-02
    99/100 stars
      Buy from Supplier

    99
    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: 99/100, based on 22 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 99 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    ecl prime western blotting detection reagent kit - by Bioz Stars, 2020-02
    99/100 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