Structured Review

GE Healthcare storm 860 phosphorimager
SUP17 alleles are aminoacylated with serine. (A) A total of 150 nM of radiolabeled tS, tS(UGG), tS(UGG) G9A , or tS(UGG) G26A were incubated with 10 μM SerRS, 10 mM serine, and 3 μM unlabeled tS. Two microliters of each reaction was treated with nuclease P1. Reaction mixtures were then spotted on glass TLC plates and chromatographed in 0.1 M ammonium acetate and 5% acetic acid. TLC plates were imaged using a <t>Storm</t> 860 <t>Phosphorimager</t> to detect free 32 P-labeled AMP and 32 P-labeled AMP charged with serine. (B) Percent of tRNA that was aminoacylated after 40 min was calculated for two technical replicates. Error bars represent the maximum and minimum values. SerRS, seryl-tRNA synthetase; tS, tRNA Ser .
Storm 860 Phosphorimager, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 96/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Evolving Mistranslating tRNAs Through a Phenotypically Ambivalent Intermediate in Saccharomyces cerevisiae"

Article Title: Evolving Mistranslating tRNAs Through a Phenotypically Ambivalent Intermediate in Saccharomyces cerevisiae

Journal: Genetics

doi: 10.1534/genetics.117.203232

SUP17 alleles are aminoacylated with serine. (A) A total of 150 nM of radiolabeled tS, tS(UGG), tS(UGG) G9A , or tS(UGG) G26A were incubated with 10 μM SerRS, 10 mM serine, and 3 μM unlabeled tS. Two microliters of each reaction was treated with nuclease P1. Reaction mixtures were then spotted on glass TLC plates and chromatographed in 0.1 M ammonium acetate and 5% acetic acid. TLC plates were imaged using a Storm 860 Phosphorimager to detect free 32 P-labeled AMP and 32 P-labeled AMP charged with serine. (B) Percent of tRNA that was aminoacylated after 40 min was calculated for two technical replicates. Error bars represent the maximum and minimum values. SerRS, seryl-tRNA synthetase; tS, tRNA Ser .
Figure Legend Snippet: SUP17 alleles are aminoacylated with serine. (A) A total of 150 nM of radiolabeled tS, tS(UGG), tS(UGG) G9A , or tS(UGG) G26A were incubated with 10 μM SerRS, 10 mM serine, and 3 μM unlabeled tS. Two microliters of each reaction was treated with nuclease P1. Reaction mixtures were then spotted on glass TLC plates and chromatographed in 0.1 M ammonium acetate and 5% acetic acid. TLC plates were imaged using a Storm 860 Phosphorimager to detect free 32 P-labeled AMP and 32 P-labeled AMP charged with serine. (B) Percent of tRNA that was aminoacylated after 40 min was calculated for two technical replicates. Error bars represent the maximum and minimum values. SerRS, seryl-tRNA synthetase; tS, tRNA Ser .

Techniques Used: Incubation, Thin Layer Chromatography, Labeling

Related Articles

Clone Assay:

Article Title: Class B scavenger receptors SR-BI/BII and CD36 mediate bacterial recognition and pro-inflammatory signaling induced by E. coli, lipopolysaccharide and cytosolic chaperonin 60
Article Snippet: The total RNA was isolated from wild type and SR-B-overexpressing clones of HeLa and HEK293 cells or from differentiated mouse BMC using the TRIzol reagent (Invitrogen) according to the manufacturer's recommended protocol. .. PCR products were separated by PAGE using 6% TBE gels (Invitrogen), and the intensity of the fluorescent signal for each band was detected using a Storm 860 PhosphorImager (GE Healthcare).

Article Title: Herbivore-Induced Defense Response in a Model Legume. Two-Spotted Spider Mites Induce Emission of (E)-β-Ocimene and Transcript Accumulation of (E)-β-Ocimene Synthase in Lotus japonicus 1
Article Snippet: The cDNA clones for LjEβOS and LjSqS were 32 P-labeled by random priming (Strip-EZ DNA, Ambion). .. The hybridized, labeled cDNA signals were detected using a Storm 860 phosphorimager (Amersham Biosciences).

Amplification:

Article Title: Class B scavenger receptors SR-BI/BII and CD36 mediate bacterial recognition and pro-inflammatory signaling induced by E. coli, lipopolysaccharide and cytosolic chaperonin 60
Article Snippet: 5′-Fluorescein-labeled primers were obtained from Operon Biotechnologies (Huntsville, AL). cDNA was amplified using System 2400 DNA thermal cycler (PerkinElmer Life Sciences) with 25, 30 or 35 cycles for GAPDH, SRBs, and the TLRs’ primers, respectively, and annealing temperatures 50°C (for GAPDH, SRBs) and 55°C (for TLRs). .. PCR products were separated by PAGE using 6% TBE gels (Invitrogen), and the intensity of the fluorescent signal for each band was detected using a Storm 860 PhosphorImager (GE Healthcare).

Electrophoresis:

Article Title: Herbivore-Induced Defense Response in a Model Legume. Two-Spotted Spider Mites Induce Emission of (E)-β-Ocimene and Transcript Accumulation of (E)-β-Ocimene Synthase in Lotus japonicus 1
Article Snippet: RNA samples (10 μ g) were isolated from L. japonicus shoots and separated by electrophoresis through formaldehyde-agarose gels and blotted to nylon membranes (Hybond-N+, Amersham Biosciences, Piscataway, NJ). .. The hybridized, labeled cDNA signals were detected using a Storm 860 phosphorimager (Amersham Biosciences).

Article Title: Factor VIII Lacking the C2 Domain Retains Cofactor Activity in Vitro *
Article Snippet: FVIII proteins or FVIIIa proteins (activated by 20 n m thrombin for 30 min at 23 °C) (0.34 μg) were subjected to electrophoresis under reducing (0.1 m dithiothreitol) conditions using 10% polyacrylamide gels at constant voltage (150 V). .. Proteins were transferred to a polyvinylidene fluoride membrane, probed with an anti-A3 monoclonal antibody (2D2), and protein bands were visualized by chemifluorescence (570 nm) using a Storm 860 phosphorimager (GE Healthcare, Piscataway, NJ).

Article Title: Mago Nashi Is Essential for Spermatogenesis in Marsilea
Article Snippet: The eluted fusion protein was then digested with 1 U of thrombin (Promega, Madison, WI) at room temperature for 16 h. Digested protein samples were subjected to preparative electrophoresis on 12.5% polyacrylamide-SDS gels. .. The antiserum was tested on immunoblots with protein isolates from microspores 4 h into development by using ECF chemiluminescence (GE Healthcare), with detection on a STORM 860 PhosphorImager (GE Healthcare) at a dilution of 1:50 ( ; ).

Quantitation Assay:

Article Title: Transactivation of PDGFR? by dopamine D4 receptor does not require PDGFR? dimerization
Article Snippet: Data Analysis Densitometry was performed on Storm 860 phosphorimager (Amersham) with ECL Plus chemiluminescent reagent (Amersham). .. Quantitation was done using ImageQuant 5.0 software (Molecular Dynamics).

Stripping Membranes:

Article Title: Herbivore-Induced Defense Response in a Model Legume. Two-Spotted Spider Mites Induce Emission of (E)-β-Ocimene and Transcript Accumulation of (E)-β-Ocimene Synthase in Lotus japonicus 1
Article Snippet: The cDNA clones for LjEβOS and LjSqS were 32 P-labeled by random priming (Strip-EZ DNA, Ambion). .. The hybridized, labeled cDNA signals were detected using a Storm 860 phosphorimager (Amersham Biosciences).

Activity Assay:

Article Title: REXO2 Is an Oligoribonuclease Active in Human Mitochondria
Article Snippet: Paragraph title: OligoRNase Activity Assays ... Immediately after the run, fluorescent products were detected using the STORM 860 PhosphorImager (GE Healthcare Life Sciences) and densitometric analysis was made with Image Quant software (GE Healthcare Life Sciences).

Resection Assay:

Article Title: PHF11 promotes DSB resection, ATR signaling, and HR
Article Snippet: Paragraph title: DNA resection assay ... Products were separated using a 1% (w/v) agarose gel run in TAE (40 mM Tris acetate at pH 8.2, 1 mM EDTA) at 400 V/h and were quantified using an Amersham Biosciences Storm 860 PhosphorImager with ImageQuaNT software (version 5.2).

BIA-KA:

Article Title: Glucagon-like peptide-2-stimulated protein synthesis through the PI 3-kinase-dependent Akt-mTOR signaling pathway
Article Snippet: Protein concentration was determined by BCA protein assay (Pierce) using bovine serum albumin as a standard. .. Western blotting images were scanned and analyzed on a Storm 860 PhosphorImager (GE Healthcare, Fairfield, CT).

Western Blot:

Article Title: Glucagon-like peptide-2-stimulated protein synthesis through the PI 3-kinase-dependent Akt-mTOR signaling pathway
Article Snippet: .. Western blotting images were scanned and analyzed on a Storm 860 PhosphorImager (GE Healthcare, Fairfield, CT). ..

Article Title: Phosducin-like protein acts as a molecular chaperone for G protein ?? dimer assembly
Article Snippet: .. Gels and immunoblots were visualized with a Storm 860 phosphorimager, and the band intensities were quantified using Image Quant software (GE Healthcare). ..

Article Title: Factor VIII Lacking the C2 Domain Retains Cofactor Activity in Vitro *
Article Snippet: Paragraph title: SDS-PAGE and Western Blotting ... Proteins were transferred to a polyvinylidene fluoride membrane, probed with an anti-A3 monoclonal antibody (2D2), and protein bands were visualized by chemifluorescence (570 nm) using a Storm 860 phosphorimager (GE Healthcare, Piscataway, NJ).

Article Title: Mago Nashi Is Essential for Spermatogenesis in Marsilea
Article Snippet: .. The antiserum was tested on immunoblots with protein isolates from microspores 4 h into development by using ECF chemiluminescence (GE Healthcare), with detection on a STORM 860 PhosphorImager (GE Healthcare) at a dilution of 1:50 ( ; ). .. For immunoblot analyses of gametophyte polypeptides, soluble proteins were isolated from identical populations of growing gametophytes at regular time intervals by using a Dounce homogenizer to fracture the spore walls.

Southern Blot:

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Article Snippet: Paragraph title: Southern Blot Analysis ... Digital images were acquired using a Storm 860 PhosphorImager and radioactivity was quantitated using ImageQuant software (Amersham Biosciences).

Northern Blot:

Article Title: Herbivore-Induced Defense Response in a Model Legume. Two-Spotted Spider Mites Induce Emission of (E)-β-Ocimene and Transcript Accumulation of (E)-β-Ocimene Synthase in Lotus japonicus 1
Article Snippet: Paragraph title: Northern Analysis ... The hybridized, labeled cDNA signals were detected using a Storm 860 phosphorimager (Amersham Biosciences).

Protein Concentration:

Article Title: Glucagon-like peptide-2-stimulated protein synthesis through the PI 3-kinase-dependent Akt-mTOR signaling pathway
Article Snippet: Protein concentration was determined by BCA protein assay (Pierce) using bovine serum albumin as a standard. .. Western blotting images were scanned and analyzed on a Storm 860 PhosphorImager (GE Healthcare, Fairfield, CT).

Polymerase Chain Reaction:

Article Title: Class B scavenger receptors SR-BI/BII and CD36 mediate bacterial recognition and pro-inflammatory signaling induced by E. coli, lipopolysaccharide and cytosolic chaperonin 60
Article Snippet: .. PCR products were separated by PAGE using 6% TBE gels (Invitrogen), and the intensity of the fluorescent signal for each band was detected using a Storm 860 PhosphorImager (GE Healthcare). .. The IL-8 secretion by HEK293 cells was analyzed in culture supernatants after a 20-h incubation in serum-free medium with or without BSA (2 mg/ml) utilizing a commercial enzyme-linked immunosorbent assay kit for human IL-8 (Invitrogen).

Article Title: Splenectomy Promotes Macrophage Polarization in a Mouse Model of Concanavalin A- (ConA-) Induced Liver Fibrosis
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Aminoacylation Assay:

Article Title: Evolving Mistranslating tRNAs Through a Phenotypically Ambivalent Intermediate in Saccharomyces cerevisiae
Article Snippet: Paragraph title: Aminoacylation assay ... The TLC plates were exposed to a phosphor screen and imaged using a Storm 860 Phosphorimager (GE Healthcare Life Sciences).

Affinity Purification:

Article Title: Mago Nashi Is Essential for Spermatogenesis in Marsilea
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Recombinant:

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Nucleic Acid Electrophoresis:

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Radioactivity:

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Isolation:

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Article Title: Herbivore-Induced Defense Response in a Model Legume. Two-Spotted Spider Mites Induce Emission of (E)-β-Ocimene and Transcript Accumulation of (E)-β-Ocimene Synthase in Lotus japonicus 1
Article Snippet: RNA samples (10 μ g) were isolated from L. japonicus shoots and separated by electrophoresis through formaldehyde-agarose gels and blotted to nylon membranes (Hybond-N+, Amersham Biosciences, Piscataway, NJ). .. The hybridized, labeled cDNA signals were detected using a Storm 860 phosphorimager (Amersham Biosciences).

Article Title: Mago Nashi Is Essential for Spermatogenesis in Marsilea
Article Snippet: The fusion protein was isolated using glutathione agarose as described by . .. The antiserum was tested on immunoblots with protein isolates from microspores 4 h into development by using ECF chemiluminescence (GE Healthcare), with detection on a STORM 860 PhosphorImager (GE Healthcare) at a dilution of 1:50 ( ; ).

Labeling:

Article Title: Herbivore-Induced Defense Response in a Model Legume. Two-Spotted Spider Mites Induce Emission of (E)-β-Ocimene and Transcript Accumulation of (E)-β-Ocimene Synthase in Lotus japonicus 1
Article Snippet: .. The hybridized, labeled cDNA signals were detected using a Storm 860 phosphorimager (Amersham Biosciences). .. Ribosomal RNA stained with ethidium bromide was visualized (ChemiImager 5500 with AlphaEaseFC software, Alpha Innotech, San Leandro, CA).

Article Title: Splenectomy Promotes Macrophage Polarization in a Mouse Model of Concanavalin A- (ConA-) Induced Liver Fibrosis
Article Snippet: Subsequently, DNA probes were created by labeling the forward primers and unlabeling the reverse primers via PCR. .. Subsequently, the screen was scanned using a storm 860 PhosphorImager, and the data were analyzed using ImageQuant software (Molecular Dynamics/GE Healthcare, Amersham, UK).

Purification:

Article Title: Mago Nashi Is Essential for Spermatogenesis in Marsilea
Article Snippet: The antiserum was batch affinity purified using purified Mv -mago protein that had been bound to the glutathione agarose beads ( ). .. The antiserum was tested on immunoblots with protein isolates from microspores 4 h into development by using ECF chemiluminescence (GE Healthcare), with detection on a STORM 860 PhosphorImager (GE Healthcare) at a dilution of 1:50 ( ; ).

Article Title: REXO2 Is an Oligoribonuclease Active in Human Mitochondria
Article Snippet: 100 fmol of single-stranded 5-mer RNA (5′-GAUCG-3′, labelled at 5′ end with Alexa 647®1005 fluorophore; Eurogentec) was incubated with either 50–500 fmol of purified recombinant REXO2 or 0.2–5 µg of total cell or mitochondrial lysates. .. Immediately after the run, fluorescent products were detected using the STORM 860 PhosphorImager (GE Healthcare Life Sciences) and densitometric analysis was made with Image Quant software (GE Healthcare Life Sciences).

Electrophoretic Mobility Shift Assay:

Article Title: Splenectomy Promotes Macrophage Polarization in a Mouse Model of Concanavalin A- (ConA-) Induced Liver Fibrosis
Article Snippet: Paragraph title: 2.9. Electrophoretic Mobility Shift Assay (EMSA) ... Subsequently, the screen was scanned using a storm 860 PhosphorImager, and the data were analyzed using ImageQuant software (Molecular Dynamics/GE Healthcare, Amersham, UK).

Polyacrylamide Gel Electrophoresis:

Article Title: Class B scavenger receptors SR-BI/BII and CD36 mediate bacterial recognition and pro-inflammatory signaling induced by E. coli, lipopolysaccharide and cytosolic chaperonin 60
Article Snippet: .. PCR products were separated by PAGE using 6% TBE gels (Invitrogen), and the intensity of the fluorescent signal for each band was detected using a Storm 860 PhosphorImager (GE Healthcare). .. The IL-8 secretion by HEK293 cells was analyzed in culture supernatants after a 20-h incubation in serum-free medium with or without BSA (2 mg/ml) utilizing a commercial enzyme-linked immunosorbent assay kit for human IL-8 (Invitrogen).

SDS Page:

Article Title: Factor VIII Lacking the C2 Domain Retains Cofactor Activity in Vitro *
Article Snippet: Paragraph title: SDS-PAGE and Western Blotting ... Proteins were transferred to a polyvinylidene fluoride membrane, probed with an anti-A3 monoclonal antibody (2D2), and protein bands were visualized by chemifluorescence (570 nm) using a Storm 860 phosphorimager (GE Healthcare, Piscataway, NJ).

Software:

Article Title: Phosducin-like protein acts as a molecular chaperone for G protein ?? dimer assembly
Article Snippet: .. Gels and immunoblots were visualized with a Storm 860 phosphorimager, and the band intensities were quantified using Image Quant software (GE Healthcare). ..

Article Title: Transactivation of PDGFR? by dopamine D4 receptor does not require PDGFR? dimerization
Article Snippet: Data Analysis Densitometry was performed on Storm 860 phosphorimager (Amersham) with ECL Plus chemiluminescent reagent (Amersham). .. Quantitation was done using ImageQuant 5.0 software (Molecular Dynamics).

Article Title: Herbivore-Induced Defense Response in a Model Legume. Two-Spotted Spider Mites Induce Emission of (E)-β-Ocimene and Transcript Accumulation of (E)-β-Ocimene Synthase in Lotus japonicus 1
Article Snippet: The hybridized, labeled cDNA signals were detected using a Storm 860 phosphorimager (Amersham Biosciences). .. Ribosomal RNA stained with ethidium bromide was visualized (ChemiImager 5500 with AlphaEaseFC software, Alpha Innotech, San Leandro, CA).

Article Title: PHF11 promotes DSB resection, ATR signaling, and HR
Article Snippet: .. Products were separated using a 1% (w/v) agarose gel run in TAE (40 mM Tris acetate at pH 8.2, 1 mM EDTA) at 400 V/h and were quantified using an Amersham Biosciences Storm 860 PhosphorImager with ImageQuaNT software (version 5.2). ..

Article Title: Hematopoietic Immortalizing Function of the NKL-Subclass Homeobox Gene TLX1
Article Snippet: .. Digital images were acquired using a Storm 860 PhosphorImager and radioactivity was quantitated using ImageQuant software (Amersham Biosciences). .. Linker-mediated nested polymerase chain reaction (LM-PCR) was performed as previously described ( ; ) with modifications that allowed the identification of genomic regions adjacent to MSCVneo-TLX1 vector sequences in a murine background.

Article Title: REXO2 Is an Oligoribonuclease Active in Human Mitochondria
Article Snippet: .. Immediately after the run, fluorescent products were detected using the STORM 860 PhosphorImager (GE Healthcare Life Sciences) and densitometric analysis was made with Image Quant software (GE Healthcare Life Sciences). .. De Novo Mitochondrial Protein Synthesis Mitochondrial protein synthesis in cultured cells was performed as described by Chomyn (1996) after addition of emetine and pulsed with [35 S] methionine/cysteine for 2 hours with a 1 hour chase.

Article Title: Splenectomy Promotes Macrophage Polarization in a Mouse Model of Concanavalin A- (ConA-) Induced Liver Fibrosis
Article Snippet: .. Subsequently, the screen was scanned using a storm 860 PhosphorImager, and the data were analyzed using ImageQuant software (Molecular Dynamics/GE Healthcare, Amersham, UK). ..

Binding Assay:

Article Title: Splenectomy Promotes Macrophage Polarization in a Mouse Model of Concanavalin A- (ConA-) Induced Liver Fibrosis
Article Snippet: Two micrograms of nuclear protein were incubated with a biotin-labeled NF-κ B p65 binding-site DNA probe (5′-AGTTGAGGGGACTTTCCCAGGC-3′; Sigma Genosys) in buffer for 30 minutes on ice. .. Subsequently, the screen was scanned using a storm 860 PhosphorImager, and the data were analyzed using ImageQuant software (Molecular Dynamics/GE Healthcare, Amersham, UK).

Agarose Gel Electrophoresis:

Article Title: PHF11 promotes DSB resection, ATR signaling, and HR
Article Snippet: .. Products were separated using a 1% (w/v) agarose gel run in TAE (40 mM Tris acetate at pH 8.2, 1 mM EDTA) at 400 V/h and were quantified using an Amersham Biosciences Storm 860 PhosphorImager with ImageQuaNT software (version 5.2). ..

Incubation:

Article Title: Glucagon-like peptide-2-stimulated protein synthesis through the PI 3-kinase-dependent Akt-mTOR signaling pathway
Article Snippet: After washing, the membrane was incubated with anti-rabbit IgG horseradish peroxidase-conjugated secondary antibody (1:3,000; Bio-Rad Laboratories, Hercules, CA) and reacted with ECL-Plus chemiluminescent detection horseradish peroxidase reagents (Amersham Biosciences, Piscataway, NJ). .. Western blotting images were scanned and analyzed on a Storm 860 PhosphorImager (GE Healthcare, Fairfield, CT).

Article Title: Phosducin-like protein acts as a molecular chaperone for G protein ?? dimer assembly
Article Snippet: The clarified lysates were incubated with 2 μg of anti-c-myc (clone 9E10, BioMol), anti-HA (clone 3F10, Roche) or anti-Flag (clone M2, Sigma) antibodies and 25 μl of a 50% slurry of Protein A/G Plus agarose (Santa Cruz Biotechnology) as described previously ( ). .. Gels and immunoblots were visualized with a Storm 860 phosphorimager, and the band intensities were quantified using Image Quant software (GE Healthcare).

Article Title: PHF11 promotes DSB resection, ATR signaling, and HR
Article Snippet: Aliquots were added to stop/loading buffer (150 mM EDTA at pH 8.0, 2% SDS, 30% glycerol, 0.1% bromophenol blue, 2–4 mg/mL Proteinase K [Roche]) and were incubated for 10 min at 37°C. .. Products were separated using a 1% (w/v) agarose gel run in TAE (40 mM Tris acetate at pH 8.2, 1 mM EDTA) at 400 V/h and were quantified using an Amersham Biosciences Storm 860 PhosphorImager with ImageQuaNT software (version 5.2).

Article Title: REXO2 Is an Oligoribonuclease Active in Human Mitochondria
Article Snippet: Incubation was carried out at 37°C, unless otherwise indicated, for 30 mins in nuclease buffer [50 mM HEPES-KOH pH 7.4, 50 mM potassium chloride, 10 mM manganese chloride, 0.01% Triton X-100, 10% glycerol and 0.1 mM DTT]. .. Immediately after the run, fluorescent products were detected using the STORM 860 PhosphorImager (GE Healthcare Life Sciences) and densitometric analysis was made with Image Quant software (GE Healthcare Life Sciences).

Article Title: Splenectomy Promotes Macrophage Polarization in a Mouse Model of Concanavalin A- (ConA-) Induced Liver Fibrosis
Article Snippet: Two micrograms of nuclear protein were incubated with a biotin-labeled NF-κ B p65 binding-site DNA probe (5′-AGTTGAGGGGACTTTCCCAGGC-3′; Sigma Genosys) in buffer for 30 minutes on ice. .. Subsequently, the screen was scanned using a storm 860 PhosphorImager, and the data were analyzed using ImageQuant software (Molecular Dynamics/GE Healthcare, Amersham, UK).

Immunoprecipitation:

Article Title: Phosducin-like protein acts as a molecular chaperone for G protein ?? dimer assembly
Article Snippet: Paragraph title: Immunoprecipitation experiments ... Gels and immunoblots were visualized with a Storm 860 phosphorimager, and the band intensities were quantified using Image Quant software (GE Healthcare).

Thin Layer Chromatography:

Article Title: Evolving Mistranslating tRNAs Through a Phenotypically Ambivalent Intermediate in Saccharomyces cerevisiae
Article Snippet: .. The TLC plates were exposed to a phosphor screen and imaged using a Storm 860 Phosphorimager (GE Healthcare Life Sciences). ..

Staining:

Article Title: Herbivore-Induced Defense Response in a Model Legume. Two-Spotted Spider Mites Induce Emission of (E)-β-Ocimene and Transcript Accumulation of (E)-β-Ocimene Synthase in Lotus japonicus 1
Article Snippet: The hybridized, labeled cDNA signals were detected using a Storm 860 phosphorimager (Amersham Biosciences). .. Ribosomal RNA stained with ethidium bromide was visualized (ChemiImager 5500 with AlphaEaseFC software, Alpha Innotech, San Leandro, CA).

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    GE Healthcare phosphorimager screening exposure
    Representative images of coronal sections of mouse brain showing PRCP mRNA expression by radioactive in situ hybridization. Panels A-D; Brain coronal section images obtained from <t>phosphorimager</t> showing the overall brain expression patter of PRCP mRNA
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    98
    GE Healthcare phosphorimaging
    Cys-8 is dispensable for glycosylation but is unstable. A , Cys-8 was substituted with serine or glycine, and the mutant proteins were transiently expressed in NIH3T3 cells. The glycan modification of the mutants was determined by PNGase F sensitivity and analyzed by immunoblotting using anti-V5 antibody. ABCB6 was still glycosylated when Cys-8 was substituted with Ser to make a consensus N -glycosylation motif. B , K562 cells were transduced with plasmids containing IRES-GFP and wild-type ABCB6-, Walker A mutant ( mt )-, C8S-, or C26A-FLAG. Cells were sorted for GFP fluorescence by FACS and analyzed by immunoblotting using anti-FLAG antibody for the expression of ABCB6 proteins. Cysteine mutants C8S and C26A were expressed at low levels. An immunoblot from a single experiment is shown. C , serine was substituted for Cys-8 and/or Cys-26 in ABCB6, or a cysteine residue was inserted between positions 9 and 10 in the C8S mutant ( C8S/C10in ). Endo H sensitivity indicates that all cysteine mutants failed to exit the ER. Cys-10 insertion did not restore the protein expression level nor the impaired ER exit. D , K562 cells stably expressing wild-type ABCB6, C8S, or C26A were labeled with [ 35 S]Met/Cys for 5 min, washed, and chased for the indicated times. Cells were harvested, and immunoprecipitated FLAG-tagged proteins were separated by SDS-PAGE and detected by <t>phosphorimaging.</t> The amount of ABCB6 proteins was analyzed by densitometry and plotted as percentage of ABCB6 protein at 0 h for each construct. Values shown are mean with the S.D. indicated by the error bars from three independent experiments. Unless otherwise noted, all experiments were repeated at least twice. A representative image from one complete experiment is shown.
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    96
    GE Healthcare storm 860 phosphorimager
    SUP17 alleles are aminoacylated with serine. (A) A total of 150 nM of radiolabeled tS, tS(UGG), tS(UGG) G9A , or tS(UGG) G26A were incubated with 10 μM SerRS, 10 mM serine, and 3 μM unlabeled tS. Two microliters of each reaction was treated with nuclease P1. Reaction mixtures were then spotted on glass TLC plates and chromatographed in 0.1 M ammonium acetate and 5% acetic acid. TLC plates were imaged using a <t>Storm</t> 860 <t>Phosphorimager</t> to detect free 32 P-labeled AMP and 32 P-labeled AMP charged with serine. (B) Percent of tRNA that was aminoacylated after 40 min was calculated for two technical replicates. Error bars represent the maximum and minimum values. SerRS, seryl-tRNA synthetase; tS, tRNA Ser .
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    Image Search Results


    Representative images of coronal sections of mouse brain showing PRCP mRNA expression by radioactive in situ hybridization. Panels A-D; Brain coronal section images obtained from phosphorimager showing the overall brain expression patter of PRCP mRNA

    Journal: Brain research

    Article Title: PROLYL CARBOXYPEPTIDASE mRNA EXPRESSION IN THE MOUSE BRAIN

    doi: 10.1016/j.brainres.2013.10.031

    Figure Lengend Snippet: Representative images of coronal sections of mouse brain showing PRCP mRNA expression by radioactive in situ hybridization. Panels A-D; Brain coronal section images obtained from phosphorimager showing the overall brain expression patter of PRCP mRNA

    Article Snippet: Emulsion autoradiography was performed after phosphorimager screening exposure (STORM 860 II phosphorimager, GE Health Care, USA).

    Techniques: Expressing, In Situ Hybridization

    Cys-8 is dispensable for glycosylation but is unstable. A , Cys-8 was substituted with serine or glycine, and the mutant proteins were transiently expressed in NIH3T3 cells. The glycan modification of the mutants was determined by PNGase F sensitivity and analyzed by immunoblotting using anti-V5 antibody. ABCB6 was still glycosylated when Cys-8 was substituted with Ser to make a consensus N -glycosylation motif. B , K562 cells were transduced with plasmids containing IRES-GFP and wild-type ABCB6-, Walker A mutant ( mt )-, C8S-, or C26A-FLAG. Cells were sorted for GFP fluorescence by FACS and analyzed by immunoblotting using anti-FLAG antibody for the expression of ABCB6 proteins. Cysteine mutants C8S and C26A were expressed at low levels. An immunoblot from a single experiment is shown. C , serine was substituted for Cys-8 and/or Cys-26 in ABCB6, or a cysteine residue was inserted between positions 9 and 10 in the C8S mutant ( C8S/C10in ). Endo H sensitivity indicates that all cysteine mutants failed to exit the ER. Cys-10 insertion did not restore the protein expression level nor the impaired ER exit. D , K562 cells stably expressing wild-type ABCB6, C8S, or C26A were labeled with [ 35 S]Met/Cys for 5 min, washed, and chased for the indicated times. Cells were harvested, and immunoprecipitated FLAG-tagged proteins were separated by SDS-PAGE and detected by phosphorimaging. The amount of ABCB6 proteins was analyzed by densitometry and plotted as percentage of ABCB6 protein at 0 h for each construct. Values shown are mean with the S.D. indicated by the error bars from three independent experiments. Unless otherwise noted, all experiments were repeated at least twice. A representative image from one complete experiment is shown.

    Journal: The Journal of Biological Chemistry

    Article Title: Conserved Intramolecular Disulfide Bond Is Critical to Trafficking and Fate of ATP-binding Cassette (ABC) Transporters ABCB6 and Sulfonylurea Receptor 1 (SUR1)/ABCC8 *

    doi: 10.1074/jbc.M110.174516

    Figure Lengend Snippet: Cys-8 is dispensable for glycosylation but is unstable. A , Cys-8 was substituted with serine or glycine, and the mutant proteins were transiently expressed in NIH3T3 cells. The glycan modification of the mutants was determined by PNGase F sensitivity and analyzed by immunoblotting using anti-V5 antibody. ABCB6 was still glycosylated when Cys-8 was substituted with Ser to make a consensus N -glycosylation motif. B , K562 cells were transduced with plasmids containing IRES-GFP and wild-type ABCB6-, Walker A mutant ( mt )-, C8S-, or C26A-FLAG. Cells were sorted for GFP fluorescence by FACS and analyzed by immunoblotting using anti-FLAG antibody for the expression of ABCB6 proteins. Cysteine mutants C8S and C26A were expressed at low levels. An immunoblot from a single experiment is shown. C , serine was substituted for Cys-8 and/or Cys-26 in ABCB6, or a cysteine residue was inserted between positions 9 and 10 in the C8S mutant ( C8S/C10in ). Endo H sensitivity indicates that all cysteine mutants failed to exit the ER. Cys-10 insertion did not restore the protein expression level nor the impaired ER exit. D , K562 cells stably expressing wild-type ABCB6, C8S, or C26A were labeled with [ 35 S]Met/Cys for 5 min, washed, and chased for the indicated times. Cells were harvested, and immunoprecipitated FLAG-tagged proteins were separated by SDS-PAGE and detected by phosphorimaging. The amount of ABCB6 proteins was analyzed by densitometry and plotted as percentage of ABCB6 protein at 0 h for each construct. Values shown are mean with the S.D. indicated by the error bars from three independent experiments. Unless otherwise noted, all experiments were repeated at least twice. A representative image from one complete experiment is shown.

    Article Snippet: 35 S-Labeled proteins were detected by phosphorimaging (Storm 860, GE Healthcare).

    Techniques: Mutagenesis, Modification, Transduction, Fluorescence, FACS, Expressing, Stable Transfection, Labeling, Immunoprecipitation, SDS Page, Construct

    SUP17 alleles are aminoacylated with serine. (A) A total of 150 nM of radiolabeled tS, tS(UGG), tS(UGG) G9A , or tS(UGG) G26A were incubated with 10 μM SerRS, 10 mM serine, and 3 μM unlabeled tS. Two microliters of each reaction was treated with nuclease P1. Reaction mixtures were then spotted on glass TLC plates and chromatographed in 0.1 M ammonium acetate and 5% acetic acid. TLC plates were imaged using a Storm 860 Phosphorimager to detect free 32 P-labeled AMP and 32 P-labeled AMP charged with serine. (B) Percent of tRNA that was aminoacylated after 40 min was calculated for two technical replicates. Error bars represent the maximum and minimum values. SerRS, seryl-tRNA synthetase; tS, tRNA Ser .

    Journal: Genetics

    Article Title: Evolving Mistranslating tRNAs Through a Phenotypically Ambivalent Intermediate in Saccharomyces cerevisiae

    doi: 10.1534/genetics.117.203232

    Figure Lengend Snippet: SUP17 alleles are aminoacylated with serine. (A) A total of 150 nM of radiolabeled tS, tS(UGG), tS(UGG) G9A , or tS(UGG) G26A were incubated with 10 μM SerRS, 10 mM serine, and 3 μM unlabeled tS. Two microliters of each reaction was treated with nuclease P1. Reaction mixtures were then spotted on glass TLC plates and chromatographed in 0.1 M ammonium acetate and 5% acetic acid. TLC plates were imaged using a Storm 860 Phosphorimager to detect free 32 P-labeled AMP and 32 P-labeled AMP charged with serine. (B) Percent of tRNA that was aminoacylated after 40 min was calculated for two technical replicates. Error bars represent the maximum and minimum values. SerRS, seryl-tRNA synthetase; tS, tRNA Ser .

    Article Snippet: The TLC plates were exposed to a phosphor screen and imaged using a Storm 860 Phosphorimager (GE Healthcare Life Sciences).

    Techniques: Incubation, Thin Layer Chromatography, Labeling