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GE Healthcare fluorescence signals
Fluorescence Signals, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 96/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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fluorescence signals - by Bioz Stars, 2020-04
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Related Articles

Diagnostic Assay:

Article Title: A Novel Quantitative Multiplex Tissue Immunoblotting for Biomarkers Predicts a Prostate Cancer Aggressive Phenotype
Article Snippet: MTI is a method that can be used to detect multiple molecular targets in FFPE tissues, while retaining both quantitative and histomorphologic diagnostic and prognostic features ( - ). .. The fluorescence signals of biomarkers and total proteins were acquired using a Typhoon 9410 imager (GE Healthcare, Piscataway, NJ) and quantified with ImageQuant5.2 software (GE Healthcare).

Amplification:

Article Title: Caenorhabditis elegans POT-1 and POT-2 Repress Telomere Maintenance Pathways
Article Snippet: C-circle quantification The C-circle amplification assay was performed as previously described ( ) with the following modifications: (1) the 96-nucleotide oligomer control was generated with a C . elegans telomeric sequence (5′ CCCATATCACTAA(GCCTAA)12 CCTCAATTCCC 3′); (2) the DNA was resolved on an agarose gel and normalized by ethidium bromide staining, and amplified DNA was dot blotted onto a neutral nylon membrane (GE Healthcare Life Sciences) and probed with a telomeric G strand (TTAGGC)3 oligo conjugated to DIG at 37°; and (3) the membrane was washed as described for the DIG Wash and Block Buffer Set (Roche) at 37° and developed with ECF reagent (GE Life Sciences). .. Fluorescence signals were collected with a Typhoon Trio scanner (GE Life Sciences) and quantified with ImageQuant TL software (GE Life Sciences) using edge subtraction.

Filtration:

Article Title: SUMOylation by SUMO2 is implicated in the degradation of misfolded ataxin-7 via RNF4 in SCA7 models
Article Snippet: Membranes were then incubated with enhanced chemiluminescence substrate (Pierce); chemiluminescence or fluorescence signals were revealed on film (ECL, Amersham Hyperfilm) or captured with an Odyssey Imaging (Li-COR) system. .. The pellet was analyzed in a filter retardation assay (SR fraction): samples (40 µg) were boiled for 5 min in 2% SDS buffer and filtered on a BRL dot-blot filtration unit through a cellulose acetate membrane (Schleicher and Schuell, 0.2 µm pore size) equilibrated with 0.1% SDS ( ; ).

Mass Spectrometry:

Article Title: Hsp90·Cdc37 Complexes with Protein Kinases Form Cooperatively with Multiple Distinct Interaction Sites *
Article Snippet: Fluorescence signals were visualized using a Typhoon 9200 phosphor- and fluorescence imaging system (GE Healthcare) with the settings appropriate for Alexa Fluor 488. .. The presence of both proteins in the respective bands was confirmed by electrospray ionization mass spectrometry on an LTQ Orbitrap XL instrument.

Cytometry:

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: Cells were immunostained with Alexa Fluor 647-conjugated antibodies and analyzed using a BD FACSCanto flow cytometer. .. Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare).

Construct:

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: The VLP release assay used for the Gag-mEos3.2 constructs was described previously ( ). .. Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare).

SYBR Green Assay:

Article Title: Preparation of DNA and protein micro arrays on glass slides coated with an agarose film
Article Snippet: The presence of immobilized DNA was also monitored by SYBR Green I ( ). .. Quantitative estimations of the fluorescence signals were made with ArrayVission™ (Imaging Research Inc., Ontario, Canada) and KaleidaGraph™.

Microarray:

Article Title: Unique Organization of Extracellular Amylases into Amylosomes in the Resistant Starch-Utilizing Human Colonic Firmicutes Bacterium Ruminococcus bromii
Article Snippet: Paragraph title: CBM-based microarray. ... The probed slides were again washed 3 times, air dried, and scanned for fluorescence signals using a Typhoon 9400 variable-mode imager (GE Healthcare Bio-Sciences AB, Uppsala, Sweden).

Incubation:

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: Cells were incubated for 16 h prior to collection of cells as suspensions and fixation with PBS containing 4% paraformaldehyde for 30 min. .. Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare).

Article Title: A Novel Quantitative Multiplex Tissue Immunoblotting for Biomarkers Predicts a Prostate Cancer Aggressive Phenotype
Article Snippet: Total proteins collected on the blotted membranes were biotinylated, and followed by incubation with Streptavidin-linked Cy5. .. The fluorescence signals of biomarkers and total proteins were acquired using a Typhoon 9410 imager (GE Healthcare, Piscataway, NJ) and quantified with ImageQuant5.2 software (GE Healthcare).

Article Title: SUMOylation by SUMO2 is implicated in the degradation of misfolded ataxin-7 via RNF4 in SCA7 models
Article Snippet: .. Membranes were then incubated with enhanced chemiluminescence substrate (Pierce); chemiluminescence or fluorescence signals were revealed on film (ECL, Amersham Hyperfilm) or captured with an Odyssey Imaging (Li-COR) system. .. Densitometry was carried out using ImageJ software (NIH).

Article Title: Proteome Profiling in Murine Models of Multiple Sclerosis: Identification of Stage Specific Markers and Culprits for Tissue Damage
Article Snippet: After 30 min incubation at 8°C in the dark, the labeling reaction was abrogated by adding 20 nmol lysine and incubating for further 10 min. .. The fluorescence signals of the three differently CyDye-labeled protein samples were imaged using a laser scanner recording band pass filtered emission wavelengths of 520 nm (Cy2); 580 nm (Cy3) and 670 nm (Cy5) respectively (Typhoon 9400 GE Healthcare).

Article Title: Unique Organization of Extracellular Amylases into Amylosomes in the Resistant Starch-Utilizing Human Colonic Firmicutes Bacterium Ruminococcus bromii
Article Snippet: Afterward, chosen Xyn-Doc samples (3 nM in blocking buffer) were incubated with the slide at room temperature for 30 min followed by 3 washing steps (5 min each) in washing buffer (TBS–10 mM CaCl2 –0.05% Tween 20). .. The probed slides were again washed 3 times, air dried, and scanned for fluorescence signals using a Typhoon 9400 variable-mode imager (GE Healthcare Bio-Sciences AB, Uppsala, Sweden).

Article Title: Hsp90·Cdc37 Complexes with Protein Kinases Form Cooperatively with Multiple Distinct Interaction Sites *
Article Snippet: After an incubation time of 4 min, the reaction was stopped by adding 200 m m Tris/HCl, pH 8.0. .. Fluorescence signals were visualized using a Typhoon 9200 phosphor- and fluorescence imaging system (GE Healthcare) with the settings appropriate for Alexa Fluor 488.

Formalin-fixed Paraffin-Embedded:

Article Title: A Novel Quantitative Multiplex Tissue Immunoblotting for Biomarkers Predicts a Prostate Cancer Aggressive Phenotype
Article Snippet: Starting with a 5μm FFPE tissue section on a standard glass slide, proteins were transferred from the tissue sections (TMA or biopsy) onto a series of overlapped thin membranes (P-Film, 20/20 GeneSystems, Inc., Rockville, MD). .. The fluorescence signals of biomarkers and total proteins were acquired using a Typhoon 9410 imager (GE Healthcare, Piscataway, NJ) and quantified with ImageQuant5.2 software (GE Healthcare).

Cell Culture:

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: For analysis of virion incorporation of CD43, CD43/6A, ICAM-1, or ICAM-1/PCT, HeLa cells cotransfected with pNL4-3 and an expression plasmid carrying one of the membrane proteins were cultured for 16 h or 2 days, and cell and virus lysates were prepared as previously described ( ). .. Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare).

Expressing:

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: Paragraph title: Cells, transfection, infection, immunostaining, and analyses for cell surface expression, virus-like particle (VLP) release, and incorporation of membrane proteins. ... Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare).

Article Title: Clathrin binding by the adaptor Ent5 promotes late stages of clathrin coat maturation
Article Snippet: Fluorescence signals were detected on a Typhoon imaging system (Amersham Biosciences, Piscataway, NJ) chemiluminescence signals were detected on a Chemi-Doc-It system (UVP, Upland, CA). .. To determine the correction factor for the antibody against Ent5, lysates of cells expressing GFP-Ent5 were collected.

Release Assay:

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: The VLP release assay used for the Gag-mEos3.2 constructs was described previously ( ). .. Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare).

Western Blot:

Article Title: A Novel Quantitative Multiplex Tissue Immunoblotting for Biomarkers Predicts a Prostate Cancer Aggressive Phenotype
Article Snippet: The fluorescence signals of biomarkers and total proteins were acquired using a Typhoon 9410 imager (GE Healthcare, Piscataway, NJ) and quantified with ImageQuant5.2 software (GE Healthcare). .. The quality of the antibodies was checked using Western blot of prostate cancer cell lines (data not shown).

Article Title: Impact of intron removal from tRNA genes on Saccharomyces cerevisiae
Article Snippet: The plasmids were introduced into appropriate strains, their yeast lysates for western blotting were prepared by alkaline lysis , and subjected to western blotting with the anti-c-Myc antibody, 9E10 (Wako Pure Chemicals, Osaka, Japan). .. For signal detection, Cy3-conjugated anti-mouse IgG antibodies and Cy5-conjugated anti-rabbit IgG antibodies (Molecular Probes, Eugene, Oregon, USA) were used as the secondary antibodies, and fluorescence signals were read with Typhoon FLA-7000 Fluorescence Scanner (GE Healthcare).

Transformation Assay:

Article Title: A Novel Quantitative Multiplex Tissue Immunoblotting for Biomarkers Predicts a Prostate Cancer Aggressive Phenotype
Article Snippet: The fluorescence signals of biomarkers and total proteins were acquired using a Typhoon 9410 imager (GE Healthcare, Piscataway, NJ) and quantified with ImageQuant5.2 software (GE Healthcare). .. The log transformed ratio was used for downstream data analysis. provides the detailed information about the primary antibodies and dilutions used in the study.

Blocking Assay:

Article Title: Caenorhabditis elegans POT-1 and POT-2 Repress Telomere Maintenance Pathways
Article Snippet: C-circle quantification The C-circle amplification assay was performed as previously described ( ) with the following modifications: (1) the 96-nucleotide oligomer control was generated with a C . elegans telomeric sequence (5′ CCCATATCACTAA(GCCTAA)12 CCTCAATTCCC 3′); (2) the DNA was resolved on an agarose gel and normalized by ethidium bromide staining, and amplified DNA was dot blotted onto a neutral nylon membrane (GE Healthcare Life Sciences) and probed with a telomeric G strand (TTAGGC)3 oligo conjugated to DIG at 37°; and (3) the membrane was washed as described for the DIG Wash and Block Buffer Set (Roche) at 37° and developed with ECF reagent (GE Life Sciences). .. Fluorescence signals were collected with a Typhoon Trio scanner (GE Life Sciences) and quantified with ImageQuant TL software (GE Life Sciences) using edge subtraction.

Article Title: Unique Organization of Extracellular Amylases into Amylosomes in the Resistant Starch-Utilizing Human Colonic Firmicutes Bacterium Ruminococcus bromii
Article Snippet: Fluorescent staining was accomplished by adding Cy3-labeled anti-Xyn and Cy5-labeled anti-CBM (diluted 1:1,000) in blocking buffer for 30 min. .. The probed slides were again washed 3 times, air dried, and scanned for fluorescence signals using a Typhoon 9400 variable-mode imager (GE Healthcare Bio-Sciences AB, Uppsala, Sweden).

Hybridization:

Article Title: Preparation of DNA and protein micro arrays on glass slides coated with an agarose film
Article Snippet: Paragraph title: Hybridization ... Quantitative estimations of the fluorescence signals were made with ArrayVission™ (Imaging Research Inc., Ontario, Canada) and KaleidaGraph™.

Transfection:

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: Paragraph title: Cells, transfection, infection, immunostaining, and analyses for cell surface expression, virus-like particle (VLP) release, and incorporation of membrane proteins. ... Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare).

Infection:

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: Paragraph title: Cells, transfection, infection, immunostaining, and analyses for cell surface expression, virus-like particle (VLP) release, and incorporation of membrane proteins. ... Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare).

Generated:

Article Title: Caenorhabditis elegans POT-1 and POT-2 Repress Telomere Maintenance Pathways
Article Snippet: C-circle quantification The C-circle amplification assay was performed as previously described ( ) with the following modifications: (1) the 96-nucleotide oligomer control was generated with a C . elegans telomeric sequence (5′ CCCATATCACTAA(GCCTAA)12 CCTCAATTCCC 3′); (2) the DNA was resolved on an agarose gel and normalized by ethidium bromide staining, and amplified DNA was dot blotted onto a neutral nylon membrane (GE Healthcare Life Sciences) and probed with a telomeric G strand (TTAGGC)3 oligo conjugated to DIG at 37°; and (3) the membrane was washed as described for the DIG Wash and Block Buffer Set (Roche) at 37° and developed with ECF reagent (GE Life Sciences). .. Fluorescence signals were collected with a Typhoon Trio scanner (GE Life Sciences) and quantified with ImageQuant TL software (GE Life Sciences) using edge subtraction.

Imaging:

Article Title: SUMOylation by SUMO2 is implicated in the degradation of misfolded ataxin-7 via RNF4 in SCA7 models
Article Snippet: .. Membranes were then incubated with enhanced chemiluminescence substrate (Pierce); chemiluminescence or fluorescence signals were revealed on film (ECL, Amersham Hyperfilm) or captured with an Odyssey Imaging (Li-COR) system. .. Densitometry was carried out using ImageJ software (NIH).

Article Title: Clathrin binding by the adaptor Ent5 promotes late stages of clathrin coat maturation
Article Snippet: .. Fluorescence signals were detected on a Typhoon imaging system (Amersham Biosciences, Piscataway, NJ) chemiluminescence signals were detected on a Chemi-Doc-It system (UVP, Upland, CA). ..

Article Title: Hsp90·Cdc37 Complexes with Protein Kinases Form Cooperatively with Multiple Distinct Interaction Sites *
Article Snippet: .. Fluorescence signals were visualized using a Typhoon 9200 phosphor- and fluorescence imaging system (GE Healthcare) with the settings appropriate for Alexa Fluor 488. ..

Sequencing:

Article Title: Caenorhabditis elegans POT-1 and POT-2 Repress Telomere Maintenance Pathways
Article Snippet: C-circle quantification The C-circle amplification assay was performed as previously described ( ) with the following modifications: (1) the 96-nucleotide oligomer control was generated with a C . elegans telomeric sequence (5′ CCCATATCACTAA(GCCTAA)12 CCTCAATTCCC 3′); (2) the DNA was resolved on an agarose gel and normalized by ethidium bromide staining, and amplified DNA was dot blotted onto a neutral nylon membrane (GE Healthcare Life Sciences) and probed with a telomeric G strand (TTAGGC)3 oligo conjugated to DIG at 37°; and (3) the membrane was washed as described for the DIG Wash and Block Buffer Set (Roche) at 37° and developed with ECF reagent (GE Life Sciences). .. Fluorescence signals were collected with a Typhoon Trio scanner (GE Life Sciences) and quantified with ImageQuant TL software (GE Life Sciences) using edge subtraction.

Article Title: Impact of intron removal from tRNA genes on Saccharomyces cerevisiae
Article Snippet: In vivo mis-decoding assay To evaluate misdecoding of an AUG codon to Ile, a c-myc tag sequence with an AUA codon as its critical 5th Ile residue and its derivative containing AUG instead of AUA were introduced at the terminus of the EGFP ORF (see Figure ). .. For signal detection, Cy3-conjugated anti-mouse IgG antibodies and Cy5-conjugated anti-rabbit IgG antibodies (Molecular Probes, Eugene, Oregon, USA) were used as the secondary antibodies, and fluorescence signals were read with Typhoon FLA-7000 Fluorescence Scanner (GE Healthcare).

Sonication:

Article Title: SUMOylation by SUMO2 is implicated in the degradation of misfolded ataxin-7 via RNF4 in SCA7 models
Article Snippet: Extracts were sonicated (four pulses of 10 s) and centrifuged at 16,000 g for 15 min at 4°C, and supernatants were collected for immunoblotting. .. Membranes were then incubated with enhanced chemiluminescence substrate (Pierce); chemiluminescence or fluorescence signals were revealed on film (ECL, Amersham Hyperfilm) or captured with an Odyssey Imaging (Li-COR) system.

Nucleic Acid Electrophoresis:

Article Title: Proteome Profiling in Murine Models of Multiple Sclerosis: Identification of Stage Specific Markers and Culprits for Tissue Damage
Article Snippet: Paragraph title: Two-dimensional difference gel electrophoresis (2D DIGE) ... The fluorescence signals of the three differently CyDye-labeled protein samples were imaged using a laser scanner recording band pass filtered emission wavelengths of 520 nm (Cy2); 580 nm (Cy3) and 670 nm (Cy5) respectively (Typhoon 9400 GE Healthcare).

In Vivo:

Article Title: Impact of intron removal from tRNA genes on Saccharomyces cerevisiae
Article Snippet: Paragraph title: In vivo mis-decoding assay ... For signal detection, Cy3-conjugated anti-mouse IgG antibodies and Cy5-conjugated anti-rabbit IgG antibodies (Molecular Probes, Eugene, Oregon, USA) were used as the secondary antibodies, and fluorescence signals were read with Typhoon FLA-7000 Fluorescence Scanner (GE Healthcare).

Fluorescence:

Article Title: Caenorhabditis elegans POT-1 and POT-2 Repress Telomere Maintenance Pathways
Article Snippet: .. Fluorescence signals were collected with a Typhoon Trio scanner (GE Life Sciences) and quantified with ImageQuant TL software (GE Life Sciences) using edge subtraction. .. POT-1 and POT-2 are negative regulators of telomere extension in vivo We obtained strains harboring the deletions pot-1 (tm1620 ) or pot-2 (tm1400 ) from Shohei Mitani and verified the presence of homozygous deletions in these strains using the polymerase chain reaction.

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: .. Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare). .. Maintenance, fixation, and immunostaining of P2 cells and their parental A3.01 cells were performed as described previously ( , ).

Article Title: A Novel Quantitative Multiplex Tissue Immunoblotting for Biomarkers Predicts a Prostate Cancer Aggressive Phenotype
Article Snippet: .. The fluorescence signals of biomarkers and total proteins were acquired using a Typhoon 9410 imager (GE Healthcare, Piscataway, NJ) and quantified with ImageQuant5.2 software (GE Healthcare). ..

Article Title: SUMOylation by SUMO2 is implicated in the degradation of misfolded ataxin-7 via RNF4 in SCA7 models
Article Snippet: .. Membranes were then incubated with enhanced chemiluminescence substrate (Pierce); chemiluminescence or fluorescence signals were revealed on film (ECL, Amersham Hyperfilm) or captured with an Odyssey Imaging (Li-COR) system. .. Densitometry was carried out using ImageJ software (NIH).

Article Title: Proteome Profiling in Murine Models of Multiple Sclerosis: Identification of Stage Specific Markers and Culprits for Tissue Damage
Article Snippet: .. The fluorescence signals of the three differently CyDye-labeled protein samples were imaged using a laser scanner recording band pass filtered emission wavelengths of 520 nm (Cy2); 580 nm (Cy3) and 670 nm (Cy5) respectively (Typhoon 9400 GE Healthcare). .. For comparison of wild type mice with or without EAE a set of 2 gels were run in a dye swap manner.

Article Title: Impact of intron removal from tRNA genes on Saccharomyces cerevisiae
Article Snippet: .. For signal detection, Cy3-conjugated anti-mouse IgG antibodies and Cy5-conjugated anti-rabbit IgG antibodies (Molecular Probes, Eugene, Oregon, USA) were used as the secondary antibodies, and fluorescence signals were read with Typhoon FLA-7000 Fluorescence Scanner (GE Healthcare). ..

Article Title: Proteomic characterization of peroxisome proliferator‐activated receptor‐γ (PPARγ) overexpressing or silenced colorectal cancer cells unveils a novel protein network associated with an aggressive phenotype), Proteomic characterization of peroxisome proliferator-activated receptor-γ (PPARγ) overexpressing or silenced colorectal cancer cells unveils a novel protein network associated with an aggressive phenotype
Article Snippet: .. Fluorescence signals were imaged by a Typhoon TRIO™ laser densitometer (GE Healthcare), recording band‐pass‐filtered emission wavelengths of 520 nm (Cy2), 580 nm (Cy3) and 670 nm (Cy5) using 100 μm as pixel size and visualized by ImageQuantTL™ software (GE Healthcare). ..

Article Title: Preparation of DNA and protein micro arrays on glass slides coated with an agarose film
Article Snippet: .. Quantitative estimations of the fluorescence signals were made with ArrayVission™ (Imaging Research Inc., Ontario, Canada) and KaleidaGraph™. .. To study the efficiency of hybridization on agarose film-coated slides we immobilized oligonucleotides containing different linkers between the 5′ end of the specific sequence ( n = 25) and the terminal primary amino group required for covalent attachment.

Article Title: Clathrin binding by the adaptor Ent5 promotes late stages of clathrin coat maturation
Article Snippet: .. Fluorescence signals were detected on a Typhoon imaging system (Amersham Biosciences, Piscataway, NJ) chemiluminescence signals were detected on a Chemi-Doc-It system (UVP, Upland, CA). ..

Article Title: Analysis of Photoreceptor Outer Segment Phagocytosis by RPE Cells in Culture
Article Snippet: .. Quantify the intensity of fluorescence signals in representative areas of each well using ImageQuant™ TL software (GE Healthcare). ..

Article Title: Unique Organization of Extracellular Amylases into Amylosomes in the Resistant Starch-Utilizing Human Colonic Firmicutes Bacterium Ruminococcus bromii
Article Snippet: .. The probed slides were again washed 3 times, air dried, and scanned for fluorescence signals using a Typhoon 9400 variable-mode imager (GE Healthcare Bio-Sciences AB, Uppsala, Sweden). ..

Article Title: Hsp90·Cdc37 Complexes with Protein Kinases Form Cooperatively with Multiple Distinct Interaction Sites *
Article Snippet: .. Fluorescence signals were visualized using a Typhoon 9200 phosphor- and fluorescence imaging system (GE Healthcare) with the settings appropriate for Alexa Fluor 488. ..

Mutagenesis:

Article Title: Impact of intron removal from tRNA genes on Saccharomyces cerevisiae
Article Snippet: Another ds oligoDNA encoding the mutant (c-Myc Ile5Met; ) was also prepared similarly. .. For signal detection, Cy3-conjugated anti-mouse IgG antibodies and Cy5-conjugated anti-rabbit IgG antibodies (Molecular Probes, Eugene, Oregon, USA) were used as the secondary antibodies, and fluorescence signals were read with Typhoon FLA-7000 Fluorescence Scanner (GE Healthcare).

Flow Cytometry:

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: Cells were immunostained with Alexa Fluor 647-conjugated antibodies and analyzed using a BD FACSCanto flow cytometer. .. Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare).

Labeling:

Article Title: Proteome Profiling in Murine Models of Multiple Sclerosis: Identification of Stage Specific Markers and Culprits for Tissue Damage
Article Snippet: Labeled samples were cup-loaded near the anodic end, and isoelectric focusing was carried out for a total of 56.000 Vh (1 h 150 V, ramp for 3 h to 300 V, ramp for 6 h to 1000 V, ramp for 3 h to 8000 V, hold at 8000 V for 4∶40 h). .. The fluorescence signals of the three differently CyDye-labeled protein samples were imaged using a laser scanner recording band pass filtered emission wavelengths of 520 nm (Cy2); 580 nm (Cy3) and 670 nm (Cy5) respectively (Typhoon 9400 GE Healthcare).

Article Title: Proteomic characterization of peroxisome proliferator‐activated receptor‐γ (PPARγ) overexpressing or silenced colorectal cancer cells unveils a novel protein network associated with an aggressive phenotype), Proteomic characterization of peroxisome proliferator-activated receptor-γ (PPARγ) overexpressing or silenced colorectal cancer cells unveils a novel protein network associated with an aggressive phenotype
Article Snippet: The experimental design using the three‐dye approach is illustrated in , and dye‐swapping among protein samples was performed to avoid artifacts due to preferential labeling. .. Fluorescence signals were imaged by a Typhoon TRIO™ laser densitometer (GE Healthcare), recording band‐pass‐filtered emission wavelengths of 520 nm (Cy2), 580 nm (Cy3) and 670 nm (Cy5) using 100 μm as pixel size and visualized by ImageQuantTL™ software (GE Healthcare).

Article Title: Hsp90·Cdc37 Complexes with Protein Kinases Form Cooperatively with Multiple Distinct Interaction Sites *
Article Snippet: 250 n m labeled hCdc37 and 500 n m labeled CeCdc37 were preincubated alone or in the presence of 2 μ m B-Raf in 20 m m Tris/HCl, pH 7.5, 50 m m KCl, 0.5 m m EDTA, 5 m m MgCl2 , 1 m m DTT, and 1% (v/v) glycerol for 10 min at 25 °C. .. Fluorescence signals were visualized using a Typhoon 9200 phosphor- and fluorescence imaging system (GE Healthcare) with the settings appropriate for Alexa Fluor 488.

Mouse Assay:

Article Title: Proteome Profiling in Murine Models of Multiple Sclerosis: Identification of Stage Specific Markers and Culprits for Tissue Damage
Article Snippet: The fluorescence signals of the three differently CyDye-labeled protein samples were imaged using a laser scanner recording band pass filtered emission wavelengths of 520 nm (Cy2); 580 nm (Cy3) and 670 nm (Cy5) respectively (Typhoon 9400 GE Healthcare). .. For comparison of wild type mice with or without EAE a set of 2 gels were run in a dye swap manner.

Dot Blot:

Article Title: SUMOylation by SUMO2 is implicated in the degradation of misfolded ataxin-7 via RNF4 in SCA7 models
Article Snippet: Membranes were then incubated with enhanced chemiluminescence substrate (Pierce); chemiluminescence or fluorescence signals were revealed on film (ECL, Amersham Hyperfilm) or captured with an Odyssey Imaging (Li-COR) system. .. The pellet was analyzed in a filter retardation assay (SR fraction): samples (40 µg) were boiled for 5 min in 2% SDS buffer and filtered on a BRL dot-blot filtration unit through a cellulose acetate membrane (Schleicher and Schuell, 0.2 µm pore size) equilibrated with 0.1% SDS ( ; ).

Immunostaining:

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: Paragraph title: Cells, transfection, infection, immunostaining, and analyses for cell surface expression, virus-like particle (VLP) release, and incorporation of membrane proteins. ... Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare).

Staining:

Article Title: Caenorhabditis elegans POT-1 and POT-2 Repress Telomere Maintenance Pathways
Article Snippet: C-circle quantification The C-circle amplification assay was performed as previously described ( ) with the following modifications: (1) the 96-nucleotide oligomer control was generated with a C . elegans telomeric sequence (5′ CCCATATCACTAA(GCCTAA)12 CCTCAATTCCC 3′); (2) the DNA was resolved on an agarose gel and normalized by ethidium bromide staining, and amplified DNA was dot blotted onto a neutral nylon membrane (GE Healthcare Life Sciences) and probed with a telomeric G strand (TTAGGC)3 oligo conjugated to DIG at 37°; and (3) the membrane was washed as described for the DIG Wash and Block Buffer Set (Roche) at 37° and developed with ECF reagent (GE Life Sciences). .. Fluorescence signals were collected with a Typhoon Trio scanner (GE Life Sciences) and quantified with ImageQuant TL software (GE Life Sciences) using edge subtraction.

Article Title: SUMOylation by SUMO2 is implicated in the degradation of misfolded ataxin-7 via RNF4 in SCA7 models
Article Snippet: Samples containing 25 µg (HEK293, HeLa cell extracts) or 50 µg (mouse brain extracts) of protein were resolved on pre-cast 4-12% (Invitrogen), 7.5% or 4-20% (Bio-Rad) gels, transferred onto nitrocellulose membranes (Protran, Whatman) by liquid transfer for 1.5 h, stained with Ponceau Red and blocked in 5% non-fat milk. .. Membranes were then incubated with enhanced chemiluminescence substrate (Pierce); chemiluminescence or fluorescence signals were revealed on film (ECL, Amersham Hyperfilm) or captured with an Odyssey Imaging (Li-COR) system.

Article Title: Unique Organization of Extracellular Amylases into Amylosomes in the Resistant Starch-Utilizing Human Colonic Firmicutes Bacterium Ruminococcus bromii
Article Snippet: Fluorescent staining was accomplished by adding Cy3-labeled anti-Xyn and Cy5-labeled anti-CBM (diluted 1:1,000) in blocking buffer for 30 min. .. The probed slides were again washed 3 times, air dried, and scanned for fluorescence signals using a Typhoon 9400 variable-mode imager (GE Healthcare Bio-Sciences AB, Uppsala, Sweden).

SDS Page:

Article Title: Proteome Profiling in Murine Models of Multiple Sclerosis: Identification of Stage Specific Markers and Culprits for Tissue Damage
Article Snippet: Second dimension SDS-PAGE was performed with homogeneous 11% polyacrylamide gels (254 by 200 mm) by the method of Tastet et al. with 150 g Tris/0.6 M HCl as gel buffer and taurine instead of glycine as buffering ion in the running buffer at 4 W/gel overnight at 25°C. .. The fluorescence signals of the three differently CyDye-labeled protein samples were imaged using a laser scanner recording band pass filtered emission wavelengths of 520 nm (Cy2); 580 nm (Cy3) and 670 nm (Cy5) respectively (Typhoon 9400 GE Healthcare).

Article Title: Clathrin binding by the adaptor Ent5 promotes late stages of clathrin coat maturation
Article Snippet: After SDS–PAGE, samples were transferred to nitrocellulose, blocked with 4% milk in TBS-T (137 mM NaCl, 15.2 mM Tris-HCl, 4.54 mM Tris, 0.896 mM Tween 20), and then probed with primary and fluorescent secondary antibodies. .. Fluorescence signals were detected on a Typhoon imaging system (Amersham Biosciences, Piscataway, NJ) chemiluminescence signals were detected on a Chemi-Doc-It system (UVP, Upland, CA).

Plasmid Preparation:

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: For analysis of virion incorporation of CD43, CD43/6A, ICAM-1, or ICAM-1/PCT, HeLa cells cotransfected with pNL4-3 and an expression plasmid carrying one of the membrane proteins were cultured for 16 h or 2 days, and cell and virus lysates were prepared as previously described ( ). .. Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare).

Article Title: Impact of intron removal from tRNA genes on Saccharomyces cerevisiae
Article Snippet: These two oligoDNAs were inserted into the BglII/EcoRI site of the plasmid pTYSC425, which harbors an EGFP gene under the control of the ADH1 promoter, to yield pSM002 and pSM004, respectively ( ). .. For signal detection, Cy3-conjugated anti-mouse IgG antibodies and Cy5-conjugated anti-rabbit IgG antibodies (Molecular Probes, Eugene, Oregon, USA) were used as the secondary antibodies, and fluorescence signals were read with Typhoon FLA-7000 Fluorescence Scanner (GE Healthcare).

Software:

Article Title: Caenorhabditis elegans POT-1 and POT-2 Repress Telomere Maintenance Pathways
Article Snippet: .. Fluorescence signals were collected with a Typhoon Trio scanner (GE Life Sciences) and quantified with ImageQuant TL software (GE Life Sciences) using edge subtraction. .. POT-1 and POT-2 are negative regulators of telomere extension in vivo We obtained strains harboring the deletions pot-1 (tm1620 ) or pot-2 (tm1400 ) from Shohei Mitani and verified the presence of homozygous deletions in these strains using the polymerase chain reaction.

Article Title: A Novel Quantitative Multiplex Tissue Immunoblotting for Biomarkers Predicts a Prostate Cancer Aggressive Phenotype
Article Snippet: .. The fluorescence signals of biomarkers and total proteins were acquired using a Typhoon 9410 imager (GE Healthcare, Piscataway, NJ) and quantified with ImageQuant5.2 software (GE Healthcare). ..

Article Title: SUMOylation by SUMO2 is implicated in the degradation of misfolded ataxin-7 via RNF4 in SCA7 models
Article Snippet: Membranes were then incubated with enhanced chemiluminescence substrate (Pierce); chemiluminescence or fluorescence signals were revealed on film (ECL, Amersham Hyperfilm) or captured with an Odyssey Imaging (Li-COR) system. .. Densitometry was carried out using ImageJ software (NIH).

Article Title: Proteome Profiling in Murine Models of Multiple Sclerosis: Identification of Stage Specific Markers and Culprits for Tissue Damage
Article Snippet: The fluorescence signals of the three differently CyDye-labeled protein samples were imaged using a laser scanner recording band pass filtered emission wavelengths of 520 nm (Cy2); 580 nm (Cy3) and 670 nm (Cy5) respectively (Typhoon 9400 GE Healthcare). .. The gels were analyzed with the different software modules of the DeCyder differential analysis software (GE Healthcare).

Article Title: Proteomic characterization of peroxisome proliferator‐activated receptor‐γ (PPARγ) overexpressing or silenced colorectal cancer cells unveils a novel protein network associated with an aggressive phenotype), Proteomic characterization of peroxisome proliferator-activated receptor-γ (PPARγ) overexpressing or silenced colorectal cancer cells unveils a novel protein network associated with an aggressive phenotype
Article Snippet: .. Fluorescence signals were imaged by a Typhoon TRIO™ laser densitometer (GE Healthcare), recording band‐pass‐filtered emission wavelengths of 520 nm (Cy2), 580 nm (Cy3) and 670 nm (Cy5) using 100 μm as pixel size and visualized by ImageQuantTL™ software (GE Healthcare). ..

Article Title: Analysis of Photoreceptor Outer Segment Phagocytosis by RPE Cells in Culture
Article Snippet: .. Quantify the intensity of fluorescence signals in representative areas of each well using ImageQuant™ TL software (GE Healthcare). ..

Microscopy:

Article Title: Basic Motifs Target PSGL-1, CD43, and CD44 to Plasma Membrane Sites Where HIV-1 Assembles
Article Snippet: Immunoblotting was performed using anti-CD43 (1G10) or anti-ICAM-1 (EP1442Y) and anti-HIV Ig in combination with appropriate secondary antibodies, and fluorescence signals were detected using a Typhoon scanner (GE Healthcare). .. For superresolution localization microscopy analysis of T cells, A3.01 cells were infected with vesicular stomatitis virus G glycoprotein (VSV-G)-pseudotyped viruses encoding Gag derivatives as described previously ( , ).

Multiplex Assay:

Article Title: A Novel Quantitative Multiplex Tissue Immunoblotting for Biomarkers Predicts a Prostate Cancer Aggressive Phenotype
Article Snippet: Paragraph title: Multiplex tissue immunoblotting (MTI) ... The fluorescence signals of biomarkers and total proteins were acquired using a Typhoon 9410 imager (GE Healthcare, Piscataway, NJ) and quantified with ImageQuant5.2 software (GE Healthcare).

Agarose Gel Electrophoresis:

Article Title: Caenorhabditis elegans POT-1 and POT-2 Repress Telomere Maintenance Pathways
Article Snippet: C-circle quantification The C-circle amplification assay was performed as previously described ( ) with the following modifications: (1) the 96-nucleotide oligomer control was generated with a C . elegans telomeric sequence (5′ CCCATATCACTAA(GCCTAA)12 CCTCAATTCCC 3′); (2) the DNA was resolved on an agarose gel and normalized by ethidium bromide staining, and amplified DNA was dot blotted onto a neutral nylon membrane (GE Healthcare Life Sciences) and probed with a telomeric G strand (TTAGGC)3 oligo conjugated to DIG at 37°; and (3) the membrane was washed as described for the DIG Wash and Block Buffer Set (Roche) at 37° and developed with ECF reagent (GE Life Sciences). .. Fluorescence signals were collected with a Typhoon Trio scanner (GE Life Sciences) and quantified with ImageQuant TL software (GE Life Sciences) using edge subtraction.

Knock-Out:

Article Title: Proteome Profiling in Murine Models of Multiple Sclerosis: Identification of Stage Specific Markers and Culprits for Tissue Damage
Article Snippet: The fluorescence signals of the three differently CyDye-labeled protein samples were imaged using a laser scanner recording band pass filtered emission wavelengths of 520 nm (Cy2); 580 nm (Cy3) and 670 nm (Cy5) respectively (Typhoon 9400 GE Healthcare). .. For the comparison of CNTF knockout mice with wild-type controls, a set of 6 gels were run, with a total of 6 individual samples per group.

Produced:

Article Title: Proteomic characterization of peroxisome proliferator‐activated receptor‐γ (PPARγ) overexpressing or silenced colorectal cancer cells unveils a novel protein network associated with an aggressive phenotype), Proteomic characterization of peroxisome proliferator-activated receptor-γ (PPARγ) overexpressing or silenced colorectal cancer cells unveils a novel protein network associated with an aggressive phenotype
Article Snippet: Fluorescence signals were imaged by a Typhoon TRIO™ laser densitometer (GE Healthcare), recording band‐pass‐filtered emission wavelengths of 520 nm (Cy2), 580 nm (Cy3) and 670 nm (Cy5) using 100 μm as pixel size and visualized by ImageQuantTL™ software (GE Healthcare). .. False positive spots, for example, produced by dye artifacts within the gel were removed manually.

Alkaline Lysis:

Article Title: Impact of intron removal from tRNA genes on Saccharomyces cerevisiae
Article Snippet: The plasmids were introduced into appropriate strains, their yeast lysates for western blotting were prepared by alkaline lysis , and subjected to western blotting with the anti-c-Myc antibody, 9E10 (Wako Pure Chemicals, Osaka, Japan). .. For signal detection, Cy3-conjugated anti-mouse IgG antibodies and Cy5-conjugated anti-rabbit IgG antibodies (Molecular Probes, Eugene, Oregon, USA) were used as the secondary antibodies, and fluorescence signals were read with Typhoon FLA-7000 Fluorescence Scanner (GE Healthcare).

Fractionation:

Article Title: SUMOylation by SUMO2 is implicated in the degradation of misfolded ataxin-7 via RNF4 in SCA7 models
Article Snippet: Paragraph title: Cell lysate fractionation, mouse brain extracts, immunoblot and filter retardation assay ... Membranes were then incubated with enhanced chemiluminescence substrate (Pierce); chemiluminescence or fluorescence signals were revealed on film (ECL, Amersham Hyperfilm) or captured with an Odyssey Imaging (Li-COR) system.

Lysis:

Article Title: SUMOylation by SUMO2 is implicated in the degradation of misfolded ataxin-7 via RNF4 in SCA7 models
Article Snippet: Mouse brain samples (cerebella, cortex, brainstem) were ground (Tissue Lyser II, Quiagen) in ice-cold RIPA lysis buffer containing 50 mM Tris-HCl, pH 8.8, 150 mM NaCl, 1 mM EDTA, 1% Na deoxycholate, 0.1% SDS, 1% NP-40, 20 mM NEM and 250 IU/ml benzonase (Merck) supplemented with protease inhibitors (Complete, Roche). .. Membranes were then incubated with enhanced chemiluminescence substrate (Pierce); chemiluminescence or fluorescence signals were revealed on film (ECL, Amersham Hyperfilm) or captured with an Odyssey Imaging (Li-COR) system.

Electrofocusing:

Article Title: Proteome Profiling in Murine Models of Multiple Sclerosis: Identification of Stage Specific Markers and Culprits for Tissue Damage
Article Snippet: The labeled samples were combined and diluted 1.33 fold by a stock solution containing 7 M urea, 2 M thiourea, 4% CHAPS, 4% IPG-buffer 4–7, 4% DTT w/v for subsequent IEF. .. The fluorescence signals of the three differently CyDye-labeled protein samples were imaged using a laser scanner recording band pass filtered emission wavelengths of 520 nm (Cy2); 580 nm (Cy3) and 670 nm (Cy5) respectively (Typhoon 9400 GE Healthcare).

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  • 91
    GE Healthcare end labeled cy5 fluorescence signal
    Linker histone H1 binding to the H3.Y nucleosome. ( A ) Schematic representation of the H1 binding assay. ( B ) Representative gel image of the H1 binding assay. Increasing amounts of H1 (0 μM: lanes 1, 2, 8 and 9; 0.4 μM: lanes 3 and 10; 0.6 μM: lanes 4 and 11; 0.7 μM: lanes 5 and 12; 0.8 μM: lanes 6 and 13; 0.9 μM: lanes 7 and 14) were mixed with H3.3 (lanes 1–7) or H3.Y (lanes 8–14) nucleosomes (0.1 μM) in presence of Nap1 (0.3 μM). After an incubation at 37°C, the complexes were detected by non-denaturing 5% PAGE with ethidium bromide staining. ( C ) Graphical representation of the H1 binding assay. The band intensities corresponding to the H1-nucleosome complex and nucleosomes were quantitated, and the rate of H1-nucleosome complex formation was plotted against the H1 concentration. The error bars indicate standard deviations (n = 3). ( D ) Hydroxyl radical footprinting of the H1-nucleosome complexes. The H3.Y and H3.3 nucleosomes were reconstituted with the <t>5′-Cy5</t> labeled 193 base-pair 601 DNA, and were purified by non-denaturing 6% polyacrylamide gel electrophoresis. Lane 1 indicates a control experiment with naked DNA. The H3.3 (lanes 2–3) or H3.Y (lanes 4–5) nucleosomes were subjected to hydroxyl radical attack in the presence (lanes 3 and 5) or absence (lanes 2 and 4) of histone H1.2. The DNA samples were then resolved on an 8% denaturing polyacrylamide gel. The arrowhead (left) and the black bar (right) indicate the nucleosomal dyad and the footprint of H1.2, respectively. ( E ) MNase treatment assay of the H1-nucleosome complex. The H1-nucleosome complex containing the H3.3 (the complex formation rate: 77%, lanes 1, 3, 5 and 7) or H3.Y nucleosome (the complex formation rate: 49%, lanes 2, 4, 6 and 8) was treated with MNase for 0, 3, 6 and 9 min for 37°C. After the incubation, the reactions were stopped by adding the proteinase K solution, containing SDS and EDTA. The DNAs were extracted, and were analyzed by non-denaturing 6% PAGE with ethidium bromide staining. ( F ) Representative gel image of the H1 binding assay with the H3.3 R42K and H3.Y K42R nucleosomes. Increasing amounts of H1 (0 μM: lanes 1, 6, 11 and 16; 0.3 μM: lanes 2, 7, 12 and 17; 0.45 μM: lanes 3, 8, 13 and 18; 0.55 μM: lanes 4, 9, 14 and 19; 0.65 μM: lanes 5, 10, 15 and 20) were mixed with H3.3 (lanes 1–4), H3.3 R42K (lanes 5–10), H3.Y (lane 11–15) or H3.Y K42R (lanes 16–20) nucleosomes (0.1 μM), in the presence of Nap1 (0.3 μM). After the incubation at 37°C, the complexes were detected by non-denaturing 5% PAGE with ethidium bromide staining. ( G ) Graphical representation of the H1 binding assay of the amino acid 42 mutants. The error bars indicate standard deviations (n = 3).
    End Labeled Cy5 Fluorescence Signal, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    GE Healthcare bodipy iam fluorescent signal intensity
    Oxidative modifications to proteins in Postoperative PCF A) Representative in-gel fluorescence image of individual patient PCF (0 and 4 hr) treated with 500 μM <t>Bodipy-iodoacetamide</t> <t>(Bodipy-IAM)</t> for 30 min to assess protein thiol modifications, where lower fluorescence intensity represents increased oxidized protein thiols. Following alkylation, 5 μg of protein was resolved by 12.5% SDS–PAGE and imaged in-gel using a Typhoon imager. The bottom panel shows coomassie blue stain to confirm equal protein loading. Quantitation of the Bodipy-IAM fluorescence signal for albumin normalized to coomassie blue protein staining. n=7 patients; Data expressed as mean ± SEM. * = p
    Bodipy Iam Fluorescent Signal Intensity, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    GE Healthcare bocillin fl fluorescence emission signal
    Activity of Bd3459 and its inhibition by ampicillin. ( A ) Pentapeptide-rich peptidoglcyan was incubated with Bd3459, Bd3459 pre-incubated with ampicillin, Bd3459 (S70A) or no enzyme, followed by digestion with cellosyl, reduction with sodium borohydride and analysis of the resulting muropeptides by HPLC. Muropeptides: 4, Tetra; 5, Penta; 44, TetraTetra; 45, TetraPenta. Peaks originating from ampicillin are indicated by a star. Bd3459, but not Bd3459 (S70A) or ampicillin-blocked Bd3459, digested Penta and TetraPenta to Tetra indicative of a DD-endo/carboxypeptidase activity. ( B ) Structures of the reduced muropeptides. GlcNAc, N-acetylglucosamine; MurNAc(r), N-acetylmuramitol; L-Ala, L-alanine; D-iGlu, D-isoglutamic acid; m -Dap, meso -diaminopimelic acid; D-Ala, D-alanine. ( C ) Bd3459 and Bd3459 (S70A) were incubated with or without ampicillin, followed by labelling with <t>Bocillin-FL</t> and SDS-polyacrylamide gel electrophoresis. Bocillin-FL was detected by fluorescence (Bocillin-FL); total protein was visualized by staining with Coomassie Blue. Bocillin-FL-binding of Bd3459 was greatly inhibited by pre-incubation with ampicillin; Bd3459 (S70A) bound substantially less Bocillin-FL than Bd3459. Thus, Bd3459 is a penicillin-binding protein and degrades isolated cell wall material.
    Bocillin Fl Fluorescence Emission Signal, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    GE Healthcare cy5 fluorescent signals
    Relative quantification of AV-derived cMyBP-C. A , A7r5 smooth muscle cells with no endogenous cMyBP-C were infected with cMyBP-C WT -AV. A7R5 cell lysates were loaded in different amounts on gel, as well as cultured cardiomyocytes infected with cMyBP-C WT or cMyBP-C C10mut . After transfer, membranes were simultaneously incubated with cMyc (rabbit polyclonal) and cMyBP-C (mouse monoclonal) antibodies. Cy3-labeled anti-mouse IgG and <t>Cy5-labeled</t> anti-rabbit IgG secondary antibodies were used to measure total cMyBP-C (endogenous + virus-derived) and cMyc (virus-derived). B , Cy3 and Cy5 samples from A7r5 samples loaded in different amounts were used to make a standard curve. These signals were used to calculate percentages of virus-derived (cMyc) signal to total cMyBP-C (cMyBP-C). C , standard curve was used to calculate percentage of virus-derived cMyBP-C to total cMyBP-C present in total lysates 48 h postinfection without virus (uninfected, UI ), with cMyBP-C WT -AV or with cMyBP-C C10mut -AV. The data are presented as means ± S.E. of three independent experiments.
    Cy5 Fluorescent Signals, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Linker histone H1 binding to the H3.Y nucleosome. ( A ) Schematic representation of the H1 binding assay. ( B ) Representative gel image of the H1 binding assay. Increasing amounts of H1 (0 μM: lanes 1, 2, 8 and 9; 0.4 μM: lanes 3 and 10; 0.6 μM: lanes 4 and 11; 0.7 μM: lanes 5 and 12; 0.8 μM: lanes 6 and 13; 0.9 μM: lanes 7 and 14) were mixed with H3.3 (lanes 1–7) or H3.Y (lanes 8–14) nucleosomes (0.1 μM) in presence of Nap1 (0.3 μM). After an incubation at 37°C, the complexes were detected by non-denaturing 5% PAGE with ethidium bromide staining. ( C ) Graphical representation of the H1 binding assay. The band intensities corresponding to the H1-nucleosome complex and nucleosomes were quantitated, and the rate of H1-nucleosome complex formation was plotted against the H1 concentration. The error bars indicate standard deviations (n = 3). ( D ) Hydroxyl radical footprinting of the H1-nucleosome complexes. The H3.Y and H3.3 nucleosomes were reconstituted with the 5′-Cy5 labeled 193 base-pair 601 DNA, and were purified by non-denaturing 6% polyacrylamide gel electrophoresis. Lane 1 indicates a control experiment with naked DNA. The H3.3 (lanes 2–3) or H3.Y (lanes 4–5) nucleosomes were subjected to hydroxyl radical attack in the presence (lanes 3 and 5) or absence (lanes 2 and 4) of histone H1.2. The DNA samples were then resolved on an 8% denaturing polyacrylamide gel. The arrowhead (left) and the black bar (right) indicate the nucleosomal dyad and the footprint of H1.2, respectively. ( E ) MNase treatment assay of the H1-nucleosome complex. The H1-nucleosome complex containing the H3.3 (the complex formation rate: 77%, lanes 1, 3, 5 and 7) or H3.Y nucleosome (the complex formation rate: 49%, lanes 2, 4, 6 and 8) was treated with MNase for 0, 3, 6 and 9 min for 37°C. After the incubation, the reactions were stopped by adding the proteinase K solution, containing SDS and EDTA. The DNAs were extracted, and were analyzed by non-denaturing 6% PAGE with ethidium bromide staining. ( F ) Representative gel image of the H1 binding assay with the H3.3 R42K and H3.Y K42R nucleosomes. Increasing amounts of H1 (0 μM: lanes 1, 6, 11 and 16; 0.3 μM: lanes 2, 7, 12 and 17; 0.45 μM: lanes 3, 8, 13 and 18; 0.55 μM: lanes 4, 9, 14 and 19; 0.65 μM: lanes 5, 10, 15 and 20) were mixed with H3.3 (lanes 1–4), H3.3 R42K (lanes 5–10), H3.Y (lane 11–15) or H3.Y K42R (lanes 16–20) nucleosomes (0.1 μM), in the presence of Nap1 (0.3 μM). After the incubation at 37°C, the complexes were detected by non-denaturing 5% PAGE with ethidium bromide staining. ( G ) Graphical representation of the H1 binding assay of the amino acid 42 mutants. The error bars indicate standard deviations (n = 3).

    Journal: Nucleic Acids Research

    Article Title: Structure and function of human histone H3.Y nucleosome

    doi: 10.1093/nar/gkw202

    Figure Lengend Snippet: Linker histone H1 binding to the H3.Y nucleosome. ( A ) Schematic representation of the H1 binding assay. ( B ) Representative gel image of the H1 binding assay. Increasing amounts of H1 (0 μM: lanes 1, 2, 8 and 9; 0.4 μM: lanes 3 and 10; 0.6 μM: lanes 4 and 11; 0.7 μM: lanes 5 and 12; 0.8 μM: lanes 6 and 13; 0.9 μM: lanes 7 and 14) were mixed with H3.3 (lanes 1–7) or H3.Y (lanes 8–14) nucleosomes (0.1 μM) in presence of Nap1 (0.3 μM). After an incubation at 37°C, the complexes were detected by non-denaturing 5% PAGE with ethidium bromide staining. ( C ) Graphical representation of the H1 binding assay. The band intensities corresponding to the H1-nucleosome complex and nucleosomes were quantitated, and the rate of H1-nucleosome complex formation was plotted against the H1 concentration. The error bars indicate standard deviations (n = 3). ( D ) Hydroxyl radical footprinting of the H1-nucleosome complexes. The H3.Y and H3.3 nucleosomes were reconstituted with the 5′-Cy5 labeled 193 base-pair 601 DNA, and were purified by non-denaturing 6% polyacrylamide gel electrophoresis. Lane 1 indicates a control experiment with naked DNA. The H3.3 (lanes 2–3) or H3.Y (lanes 4–5) nucleosomes were subjected to hydroxyl radical attack in the presence (lanes 3 and 5) or absence (lanes 2 and 4) of histone H1.2. The DNA samples were then resolved on an 8% denaturing polyacrylamide gel. The arrowhead (left) and the black bar (right) indicate the nucleosomal dyad and the footprint of H1.2, respectively. ( E ) MNase treatment assay of the H1-nucleosome complex. The H1-nucleosome complex containing the H3.3 (the complex formation rate: 77%, lanes 1, 3, 5 and 7) or H3.Y nucleosome (the complex formation rate: 49%, lanes 2, 4, 6 and 8) was treated with MNase for 0, 3, 6 and 9 min for 37°C. After the incubation, the reactions were stopped by adding the proteinase K solution, containing SDS and EDTA. The DNAs were extracted, and were analyzed by non-denaturing 6% PAGE with ethidium bromide staining. ( F ) Representative gel image of the H1 binding assay with the H3.3 R42K and H3.Y K42R nucleosomes. Increasing amounts of H1 (0 μM: lanes 1, 6, 11 and 16; 0.3 μM: lanes 2, 7, 12 and 17; 0.45 μM: lanes 3, 8, 13 and 18; 0.55 μM: lanes 4, 9, 14 and 19; 0.65 μM: lanes 5, 10, 15 and 20) were mixed with H3.3 (lanes 1–4), H3.3 R42K (lanes 5–10), H3.Y (lane 11–15) or H3.Y K42R (lanes 16–20) nucleosomes (0.1 μM), in the presence of Nap1 (0.3 μM). After the incubation at 37°C, the complexes were detected by non-denaturing 5% PAGE with ethidium bromide staining. ( G ) Graphical representation of the H1 binding assay of the amino acid 42 mutants. The error bars indicate standard deviations (n = 3).

    Article Snippet: The end labeled Cy5 fluorescence signal was detected through a glass plate on a Typhoon 9410 imager (GE Healthcare).

    Techniques: Binding Assay, Incubation, Polyacrylamide Gel Electrophoresis, Staining, Concentration Assay, Footprinting, Labeling, Purification

    Oxidative modifications to proteins in Postoperative PCF A) Representative in-gel fluorescence image of individual patient PCF (0 and 4 hr) treated with 500 μM Bodipy-iodoacetamide (Bodipy-IAM) for 30 min to assess protein thiol modifications, where lower fluorescence intensity represents increased oxidized protein thiols. Following alkylation, 5 μg of protein was resolved by 12.5% SDS–PAGE and imaged in-gel using a Typhoon imager. The bottom panel shows coomassie blue stain to confirm equal protein loading. Quantitation of the Bodipy-IAM fluorescence signal for albumin normalized to coomassie blue protein staining. n=7 patients; Data expressed as mean ± SEM. * = p

    Journal: Laboratory investigation; a journal of technical methods and pathology

    Article Title: Hemoglobin-associated Oxidative Stress in the Pericardial Compartment of Post-operative Cardiac Surgery Patients

    doi: 10.1038/labinvest.2014.144

    Figure Lengend Snippet: Oxidative modifications to proteins in Postoperative PCF A) Representative in-gel fluorescence image of individual patient PCF (0 and 4 hr) treated with 500 μM Bodipy-iodoacetamide (Bodipy-IAM) for 30 min to assess protein thiol modifications, where lower fluorescence intensity represents increased oxidized protein thiols. Following alkylation, 5 μg of protein was resolved by 12.5% SDS–PAGE and imaged in-gel using a Typhoon imager. The bottom panel shows coomassie blue stain to confirm equal protein loading. Quantitation of the Bodipy-IAM fluorescence signal for albumin normalized to coomassie blue protein staining. n=7 patients; Data expressed as mean ± SEM. * = p

    Article Snippet: The Bodipy-IAM fluorescent signal intensity for albumin was quantified using ImageQuantTL analysis software (GE Healthcare Biosciences, Pittsburgh, PA) and the coomassie blue protein stain for albumin was quantified using the AlphaView SA software.

    Techniques: Fluorescence, SDS Page, Staining, Quantitation Assay

    Activity of Bd3459 and its inhibition by ampicillin. ( A ) Pentapeptide-rich peptidoglcyan was incubated with Bd3459, Bd3459 pre-incubated with ampicillin, Bd3459 (S70A) or no enzyme, followed by digestion with cellosyl, reduction with sodium borohydride and analysis of the resulting muropeptides by HPLC. Muropeptides: 4, Tetra; 5, Penta; 44, TetraTetra; 45, TetraPenta. Peaks originating from ampicillin are indicated by a star. Bd3459, but not Bd3459 (S70A) or ampicillin-blocked Bd3459, digested Penta and TetraPenta to Tetra indicative of a DD-endo/carboxypeptidase activity. ( B ) Structures of the reduced muropeptides. GlcNAc, N-acetylglucosamine; MurNAc(r), N-acetylmuramitol; L-Ala, L-alanine; D-iGlu, D-isoglutamic acid; m -Dap, meso -diaminopimelic acid; D-Ala, D-alanine. ( C ) Bd3459 and Bd3459 (S70A) were incubated with or without ampicillin, followed by labelling with Bocillin-FL and SDS-polyacrylamide gel electrophoresis. Bocillin-FL was detected by fluorescence (Bocillin-FL); total protein was visualized by staining with Coomassie Blue. Bocillin-FL-binding of Bd3459 was greatly inhibited by pre-incubation with ampicillin; Bd3459 (S70A) bound substantially less Bocillin-FL than Bd3459. Thus, Bd3459 is a penicillin-binding protein and degrades isolated cell wall material.

    Journal: PLoS Pathogens

    Article Title: Specialized Peptidoglycan Hydrolases Sculpt the Intra-bacterial Niche of Predatory Bdellovibrio and Increase Population Fitness

    doi: 10.1371/journal.ppat.1002524

    Figure Lengend Snippet: Activity of Bd3459 and its inhibition by ampicillin. ( A ) Pentapeptide-rich peptidoglcyan was incubated with Bd3459, Bd3459 pre-incubated with ampicillin, Bd3459 (S70A) or no enzyme, followed by digestion with cellosyl, reduction with sodium borohydride and analysis of the resulting muropeptides by HPLC. Muropeptides: 4, Tetra; 5, Penta; 44, TetraTetra; 45, TetraPenta. Peaks originating from ampicillin are indicated by a star. Bd3459, but not Bd3459 (S70A) or ampicillin-blocked Bd3459, digested Penta and TetraPenta to Tetra indicative of a DD-endo/carboxypeptidase activity. ( B ) Structures of the reduced muropeptides. GlcNAc, N-acetylglucosamine; MurNAc(r), N-acetylmuramitol; L-Ala, L-alanine; D-iGlu, D-isoglutamic acid; m -Dap, meso -diaminopimelic acid; D-Ala, D-alanine. ( C ) Bd3459 and Bd3459 (S70A) were incubated with or without ampicillin, followed by labelling with Bocillin-FL and SDS-polyacrylamide gel electrophoresis. Bocillin-FL was detected by fluorescence (Bocillin-FL); total protein was visualized by staining with Coomassie Blue. Bocillin-FL-binding of Bd3459 was greatly inhibited by pre-incubation with ampicillin; Bd3459 (S70A) bound substantially less Bocillin-FL than Bd3459. Thus, Bd3459 is a penicillin-binding protein and degrades isolated cell wall material.

    Article Snippet: The Bocillin-FL fluorescence emission signal at 520 nm was scanned with a Typhoon reader (GE Healthcare, Little Chalfont, UK) upon excitation at 488 nm.

    Techniques: Activity Assay, Inhibition, Incubation, High Performance Liquid Chromatography, Polyacrylamide Gel Electrophoresis, Fluorescence, Staining, Binding Assay, Isolation

    Relative quantification of AV-derived cMyBP-C. A , A7r5 smooth muscle cells with no endogenous cMyBP-C were infected with cMyBP-C WT -AV. A7R5 cell lysates were loaded in different amounts on gel, as well as cultured cardiomyocytes infected with cMyBP-C WT or cMyBP-C C10mut . After transfer, membranes were simultaneously incubated with cMyc (rabbit polyclonal) and cMyBP-C (mouse monoclonal) antibodies. Cy3-labeled anti-mouse IgG and Cy5-labeled anti-rabbit IgG secondary antibodies were used to measure total cMyBP-C (endogenous + virus-derived) and cMyc (virus-derived). B , Cy3 and Cy5 samples from A7r5 samples loaded in different amounts were used to make a standard curve. These signals were used to calculate percentages of virus-derived (cMyc) signal to total cMyBP-C (cMyBP-C). C , standard curve was used to calculate percentage of virus-derived cMyBP-C to total cMyBP-C present in total lysates 48 h postinfection without virus (uninfected, UI ), with cMyBP-C WT -AV or with cMyBP-C C10mut -AV. The data are presented as means ± S.E. of three independent experiments.

    Journal: The Journal of Biological Chemistry

    Article Title: A Hypertrophic Cardiomyopathy-associated MYBPC3 Mutation Common in Populations of South Asian Descent Causes Contractile Dysfunction *

    doi: 10.1074/jbc.M114.607911

    Figure Lengend Snippet: Relative quantification of AV-derived cMyBP-C. A , A7r5 smooth muscle cells with no endogenous cMyBP-C were infected with cMyBP-C WT -AV. A7R5 cell lysates were loaded in different amounts on gel, as well as cultured cardiomyocytes infected with cMyBP-C WT or cMyBP-C C10mut . After transfer, membranes were simultaneously incubated with cMyc (rabbit polyclonal) and cMyBP-C (mouse monoclonal) antibodies. Cy3-labeled anti-mouse IgG and Cy5-labeled anti-rabbit IgG secondary antibodies were used to measure total cMyBP-C (endogenous + virus-derived) and cMyc (virus-derived). B , Cy3 and Cy5 samples from A7r5 samples loaded in different amounts were used to make a standard curve. These signals were used to calculate percentages of virus-derived (cMyc) signal to total cMyBP-C (cMyBP-C). C , standard curve was used to calculate percentage of virus-derived cMyBP-C to total cMyBP-C present in total lysates 48 h postinfection without virus (uninfected, UI ), with cMyBP-C WT -AV or with cMyBP-C C10mut -AV. The data are presented as means ± S.E. of three independent experiments.

    Article Snippet: Cy3 and Cy5 fluorescent signals were obtained on a Typhoon 9200 scanner (GE Healthcare Life Sciences).

    Techniques: Derivative Assay, Infection, Cell Culture, Incubation, Labeling