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Molecular dynamics phosphorimager

Manufactured by GE Healthcare
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The Molecular Dynamics Phosphorimager is a laboratory instrument designed for the detection and analysis of radioactively labeled biomolecules, such as DNA, RNA, and proteins. The core function of this product is to capture and quantify the spatial distribution and intensity of radioactive signals, enabling researchers to visualize and analyze the presence and abundance of specific molecules in their samples.

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Lab products found in correlation

Imagequant 5, by GE Healthcare (2 mentions) Amersham hybond n, by GE Healthcare (2 mentions) Amv reverse transcriptase, by Promega (1 mentions) Tnt system, by Promega (1 mentions) Imagequant software, by GE Healthcare (1 mentions) γ 32p atp, by PerkinElmer (1 mentions)

Close spelling variants of this product

Storm 860 Molecular Dynamics phosphorimager PhosphorImager

5 protocols using molecular dynamics phosphorimager

1

Quantifying Ribosome-Bound mRNA Levels

Cited 1 time
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For ribosome association of individual mRNAs, gel electrophoresis following polysome analysis and fractionation was performed as described above. RNA was subsequently transferred to nitrocellulose membranes (AmershamHybond™–N, GE Healthcare) and further processed as described 31 (link). DNA oligonucleotides (Supplementary Information Table S2 [X85, X96, X105, X114]) were radiolabeled with PNK according to standard procedures. Probes were incubated with the membranes in hybridization buffer (6 × SSC, 0.1% SDS, 10 × Denhardt’s reagent) overnight at 42°C. Membranes were subsequently washed three times with wash buffer (6 × SSC, 0.1% SDS). Probe signals were visualized by a Molecular Dynamics Phosphorimager (GE Healthcare) and quantified by ImageQuant 5.2 software (Molecular Dynamics). Normalized signal intensities were compiled for fractions corresponding to monosomes, light and heavy polysomes and averaged from at least three biological replicates.
Guenther U.P., Weinberg D.E., Zubradt M.M., Tedeschi F.A., Stawicki B.N., Zagore L.L., Brar G.A., Licatalosi D.D., Bartel D.P., Weissman J.S, & Jankowsky E. (2018). The helicase Ded1p controls use of near-cognate translation initiation codons in 5′UTRs. Nature, 559(7712), 130-134.
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2

Primer Extension Analysis of RNA

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Primer extension analysis using 5 μg of total RNA was performed as previously described (5 (link)). Annealing of the 5΄-end-labeled primer Y25 was performed in 1xRT-buffer by heating for 3 min to 80°C, snap freezing in liquid nitrogen, and slowly thawing on ice. Primer extension reactions were performed in RT-buffer by using the AMV reverse transcriptase (Promega, WI, USA) by incubation at 42°C for 30 min. The samples were separated on an 8% PAA-8 M urea gel, and the extension signals were visualized by using a Molecular Dynamics PhosphorImager (GE Healthcare, NJ, USA).
Temmel H., Müller C., Sauert M., Vesper O., Reiss A., Popow J., Martinez J, & Moll I. (2016). The RNA ligase RtcB reverses MazF-induced ribosome heterogeneity in Escherichia coli. Nucleic Acids Research, 45(8), 4708-4721.
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3

In vitro Ubiquitination Assay

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E1, E2, Cdh1, and APC/C were expressed and purified as described previously (Carroll et al., 2005 (link); Enquist-Newman et al., 2008 (link)). Substrates were transcribed and translated in vitro using the TNT system (Promega, Madison, WI) from plasmids with 35S-methionine and treated with 10 mM NEM (10 minutes) followed by 20 mM DTT (10 minutes) to inactivate ubiquitin chain-extending activities in the reticulocyte lysate. E1 (Uba1, 300 nM), E2 (Ubc4, 50 µM), ubiquitin (150 µM), and ATP (1 mM) were incubated for 15 minutes. APC/C (0.1–1 nM), substrate (2 µl of TnT mix into 15 µl reaction), and Cdh1 (2 µl of TnT mix into 15 µl reaction) were added. Reaction were incubated for the indicated times at room temperature, stopped by the addition of SDS sample buffer, separated by SDS-PAGE, and visualized and quantified with a Molecular Dynamics Phosphorimager (GE Healthcare, Fairfield, CT).
Matsusaka T., Enquist-Newman M., Morgan D.O, & Pines J. (2014). Co-activator independent differences in how the metaphase and anaphase APC/C recognise the same substrate. Biology Open, 3(10), 904-912.
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4

Measuring ATPase Activity of Chromatin Remodelers

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The ATPase activity of each remodeling enzyme was determined
(see also (Smith and Peterson,
2005)) at 30 °C using 100 μM ATP and 0.2
μCi of [γ-32P]-ATP (Perkin Elmer) in buffer A
(10 mM Tris-HCl, pH 8.0, 70 mM NaCl, 5 mM MgCl2, 0.1 mg/ml
BSA, and 1 mM DTT). 0.1 mg/ml plasmid DNA was used as a substrate.
Released phosphate was resolved from ATP by thin-layer chromatography
PEI Cellulose (Millipore) in 750 mM potassium phosphate, pH 3.5.
Analysis of hydrolysis rates was performed using a Molecular Dynamics
PhosphorImager and Image-Quant software (GE Healthcare). ATP-hydrolysis
rates were determined over three linear time points. The concentration
of each remodeling enzyme was estimated relative to a standard SWI/SNF
preparation.
Krietenstein N., Wal M., Watanabe S., Park B., Peterson C.L., Pugh B.F, & Korber P. (2016). Genomic nucleosome organization reconstituted with pure proteins. Cell, 167(3), 709-721.e12.
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5

Quantifying Ribosome-Bound mRNA Levels

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
For ribosome association of individual mRNAs, gel electrophoresis following polysome analysis and fractionation was performed as described above. RNA was subsequently transferred to nitrocellulose membranes (AmershamHybond™–N, GE Healthcare) and further processed as described 31 (link). DNA oligonucleotides (Supplementary Information Table S2 [X85, X96, X105, X114]) were radiolabeled with PNK according to standard procedures. Probes were incubated with the membranes in hybridization buffer (6 × SSC, 0.1% SDS, 10 × Denhardt’s reagent) overnight at 42°C. Membranes were subsequently washed three times with wash buffer (6 × SSC, 0.1% SDS). Probe signals were visualized by a Molecular Dynamics Phosphorimager (GE Healthcare) and quantified by ImageQuant 5.2 software (Molecular Dynamics). Normalized signal intensities were compiled for fractions corresponding to monosomes, light and heavy polysomes and averaged from at least three biological replicates.
Guenther U.P., Weinberg D.E., Zubradt M.M., Tedeschi F.A., Stawicki B.N., Zagore L.L., Brar G.A., Licatalosi D.D., Bartel D.P., Weissman J.S, & Jankowsky E. (2018). The helicase Ded1p controls use of near-cognate translation initiation codons in 5′UTRs. Nature, 559(7712), 130-134.
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