Log In Sign up
The largest database of trusted experimental protocols

Amersham typhoon ip

Manufactured by GE Healthcare
Visit Supplier

The Amersham Typhoon IP is a phosphorimager designed for the detection and quantification of radiolabeled biomolecules in a variety of samples, including gels, membranes, and microplates. It utilizes a laser-based scanning system to capture high-resolution images of radioactive signals, enabling sensitive and accurate analysis of experimental results.

Automatically generated - may contain errors

Lab products found in correlation

γ 32p atp, by PerkinElmer (3 mentions) Bas ip ms 3543, by GE Healthcare (2 mentions) T4 polynucleotide kinase, by New England Biolabs (1 mentions) Nylon membrane, by Cytiva (1 mentions) Polygram cel 300 pei uv254, by Macherey-Nagel (1 mentions)

Spelling variants of this product

Amersham Typhoon (41 citations) Amersham Typhoon IP Biomolecular Imager (3 citations)

8 protocols using amersham typhoon ip

1

Telomerase Activity and Processivity Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols
Telomerase activity assays were carried out for 1 hour at 30 °C as previously described in reaction buffer (50 mM TRIS-HCl pH 8.0, 50 mM or 150 mM KCl, 1 mM MgCl2) 49 . Assays with telomerase alone were carried out with 50 nM primer A, 50 mM KCl, 10 μM or 50 μM dATP, dTTP, and dGTP, and 0.33 μM dGTP [α−32P] (Perkin Elmer). Telomerase assays to assess POT1/TPP1 stimulation were carried out with 50 nM primer A5, 150 mM KCl, 100 nM POT1/TPP1, 500 μM dATP, dTTP, 3.3 μM dGTP, and 0.33 μM dGTP [α−32P] (Perkin Elmer). POT1/TPP1 was purified as previously described 15 . Telomerase products were separated on 12% poly-acrylamide, 1xTBE, 7 M Urea, sequencing gels and detected using storage phosphor screens (GE Healthcare Life Sciences, BAS-IP MS 3543) and an Amersham Typhoon IP (GE Healthcare Life Sciences). Phosphorylated primer A-2 was used as a loading control. Activity was quantified as total radioactive incorporated per lane normalized to the loading control. To compare activity between 3xFLAG- and 3xFLAG-HaloTag containing telomerase, specific activity was calculated by dividing the activity measured by telomerase assays by the TR levels determined by northern blot. Processivity was quantified as the intensity of products longer than 6 repeats divided by the total signal per lane.
Patrick E.M., Slivka J., Payne B., Comstock M.J, & Schmidt J.C. (2020). Observation of processive telomerase catalysis using high-resolution optical tweezers. Nature chemical biology, 16(7), 801-809.
+ Open protocol
+ Expand
2

RNA Duplex Unwinding Kinetics by eIF4A1

Check if the same lab product or an alternative is used in the 5 most similar protocols
RNA duplex was prepared by annealing of 5′-GCGUCUUUACGGUGCUUAAAACAAAACAAAACAAAACAAAA-3′ and 5′-AGCACCGUAAAGACGC-3′, and short RNA strand was radiolabeled by γ 32P-ATP (Perkinelmer) and T4 polynucleotide kinase (New England Biolabs) (Jankowsky and Putnam, 2010 (link); Floor et al., 2016 (link)). In 30 l reaction scale, 10 M recombinant eIF4A1 and 20 nM of the radio-labeled RNA duplex was incubated in 40 mM Tris-HCl pH 7.5, 2 mM HEPES-NaOH pH 7.5, 15 mM NaCl, 2 mM ATP, 2.5 mM MgCl2, 0.01% IGEPAL CA-630, 1% glycerol, 2 mM DTT, and 2 μM complementary DNA (5′-AGCACCGTAAAGACGC-3′) at 37°C. The reaction was quenched by an addition of equal amount of 2× Gel Loading Dye, Purple (New England Biolabs) and run on native TBE polyacrylamide gel, 20%. The images were obtained by Amersham Typhoon IP (GE Healthcare).
Iwasaki S., Iwasaki W., Takahashi M., Sakamoto A., Watanabe C., Shichino Y., Floor S.N., Fujiwara K., Mito M., Dodo K., Sodeoka M., Imataka H., Honma T., Fukuzawa K., Ito T, & Ingolia N.T. (2018). The Translation Inhibitor Rocaglamide Targets a Biomolecular Cavity between eIF4A and Polypurine RNA. Molecular cell, 73(4), 738-748.e9.
+ Open protocol
+ Expand
3

Detecting DNA Adducts via TLC Postlabelling

Check if the same lab product or an alternative is used in the 5 most similar protocols
The nuclease P1 enrichment version of the thin-layer chromatography (TLC) 32P-postlabelling assay was used to detect DNA adducts as reported [14 ,24 (link)]. After chromatography, TLC sheets were scanned using phosphor-imaging technology (Amersham™ Typhoon IP; GE Healthcare). Adduct levels were calculated as relative adduct labelling (RAL) values, which represent the ratio of count rates of adducted nucleotides (adducts) over count rates of total (adducted plus normal) nucleotides in the DNA sample analysed. A deoxyadenosine-3′-monophosphate (dAp) standard was labelled in each experiment to estimate the count rates of total nucleotides. Results are expressed as DNA adducts/108 nucleotides.
Reed L., Jarvis I.W., Phillips D.H, & Arlt V.M. (2020). Enhanced DNA adduct formation by benzo[a]pyrene in human liver cells lacking cytochrome P450 oxidoreductase. Mutation Research. Genetic Toxicology and Environmental Mutagenesis, 852, 503162.
+ Open protocol
+ Expand
4

In Vitro and Ex Vivo ARG of [18F]TZ-Z09591

Check if the same lab product or an alternative is used in the 5 most similar protocols
In order to study the specificity of [18F]TZ-Z09591 in vitro, ARG binding studies were performed on biopsies, and cell pellets sectioned with a cryostat microtome (Micron HM560), and mounted on glass slides. See supplemental materials for details on the protocol. The samples, optionally pre-treated with blocking compound, were incubated in a solution containing 5 nM of [18F]TZ-Z09591 (0.1 MBq/mL) for one-hour incubation at room temperature, followed by washes in cold PBS / 1% bovine serum albumin (BSA) solution and cold Milli-Q water. A reference consisting of a 10µL drop of the radiotracer-containing solution of known radioactivity was placed on an absorbent paper glued to a glass slide. The sections and reference were then placed in contact with a phosphor-imaging plate overnight. Once the exposure was complete, the plate was scanned by a phosphor-imager system (Amersham Typhoon IP, GE Healthcare). The ImageJ software (ImageJ 1.45S) was used to visualize and analyze the sections. The data obtained from in vitro ARG are expressed as fmol/mm3.
Ex vivo autoradiography follow the same protocol, except that the organs collected from animals injected with [18F]TZ-Z09591 were snap-frozen, and the sections obtain from these were directly placed in contact with a phosphor-imaging plate.
Wegrzyniak O., Zhang B., Rokka J., Rosestedt M., Mitran B., Cheung P., Puuvuori E., Ingvast S., Persson J., Nordström H., Löfblom J., Pontén F., Frejd F.Y., Korsgren O., Eriksson J, & Eriksson O. (2023). Imaging of fibrogenesis in the liver by [18F]TZ-Z09591, an Affibody molecule targeting platelet derived growth factor receptor β. EJNMMI Radiopharmacy and Chemistry, 8, 23.
+ Open protocol
+ Expand
5

Quantifying eIF4A1 ATP Binding Kinetics

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Ten μM of recombinant eIF4A1 proteins and 0.1 μM of γ 32P-ATP (Perkinelmer) were incubated for 15 min at 37°C and then UV-exposed for 1500 mJ/cm2 at 254 nm (Iwasaki et al., 2016 (link)). The complexes were separated in SDS-PAGE and the images were obtained by Amersham Typhoon IP (GE Healthcare).
Iwasaki S., Iwasaki W., Takahashi M., Sakamoto A., Watanabe C., Shichino Y., Floor S.N., Fujiwara K., Mito M., Dodo K., Sodeoka M., Imataka H., Honma T., Fukuzawa K., Ito T, & Ingolia N.T. (2018). The Translation Inhibitor Rocaglamide Targets a Biomolecular Cavity between eIF4A and Polypurine RNA. Molecular cell, 73(4), 738-748.e9.
+ Open protocol
+ Expand
6

Small RNA Northern Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Small RNA northern blot analyses were performed as previously described (Choi et al., 2020) . Total RNA was extracted from 10-day-old seedlings of WT, fha2-2, hyl1-2, and fha2-2 hyl1-2 using XENOPURE reagent (Xenohelix, Jeonju-si, Republic of Korea). The extracted RNA was precipitated twice with 2-propanol (100% and 75%) and dissolved in 50% formamide. The purified RNA was separated using 5%-15% denaturing polyacrylamide gel electrophoresis (National Diagnostics, Atlanta, GA). The resolved RNA was transferred onto a nylon membrane (Amersham, Amersham, UK). The membrane blots were incubated with endlabeled DNA probes for 12 h for hybridization (Ambion, Austin, TX). The blots were then washed twice with washing buffer (saline-sodium citrate and 0.1% sodium dodecyl sulfate, SDS) for 20 min each. Hybridization signals were detected using Amersham Typhoon IP (GE).
Park S.J., Choi S.W., Kim G.M., Møller C., Pai H.S, & Yang S.W. (2021). Light-stabilized FHA2 suppresses miRNA biogenesis through interactions with DCL1 and HYL1. Molecular plant, 14(4).
+ Open protocol
+ Expand
7

eIF4A1 ATPase Activity Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols
In 20 μl reaction scale, 10 μM recombinant eIF4A1 and 0.25 μM γ 32P-ATP (Perkinelmer) are incubated in 20 mM HEPES-NaOH pH 7.5, 150 mM NaCl, 10% glycerol, 1 mM DTT, and 1 mM MgCl2 at 37°C. The reaction was quenched by adding the equal volume of 200 mM Tris-HCl, pH 7.5, 25 mM EDTA pH 8.0, 300 mM NaCl, and 2% sodium dodecyl sulfate (SDS), spot on thin layer chromatography plate (Polygram CEL 300 PEI / UV254, Macherey-Nagel), and developed by 0.45 M NH4SO4. The images were obtained by Amersham Typhoon IP (GE Healthcare).
Iwasaki S., Iwasaki W., Takahashi M., Sakamoto A., Watanabe C., Shichino Y., Floor S.N., Fujiwara K., Mito M., Dodo K., Sodeoka M., Imataka H., Honma T., Fukuzawa K., Ito T, & Ingolia N.T. (2018). The Translation Inhibitor Rocaglamide Targets a Biomolecular Cavity between eIF4A and Polypurine RNA. Molecular cell, 73(4), 738-748.e9.
+ Open protocol
+ Expand
8

Telomerase Activity and Processivity Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols
Telomerase activity assays were carried out for 1 hour at 30 °C as previously described in reaction buffer (50 mM TRIS-HCl pH 8.0, 50 mM or 150 mM KCl, 1 mM MgCl2) 49 . Assays with telomerase alone were carried out with 50 nM primer A, 50 mM KCl, 10 μM or 50 μM dATP, dTTP, and dGTP, and 0.33 μM dGTP [α−32P] (Perkin Elmer). Telomerase assays to assess POT1/TPP1 stimulation were carried out with 50 nM primer A5, 150 mM KCl, 100 nM POT1/TPP1, 500 μM dATP, dTTP, 3.3 μM dGTP, and 0.33 μM dGTP [α−32P] (Perkin Elmer). POT1/TPP1 was purified as previously described 15 . Telomerase products were separated on 12% poly-acrylamide, 1xTBE, 7 M Urea, sequencing gels and detected using storage phosphor screens (GE Healthcare Life Sciences, BAS-IP MS 3543) and an Amersham Typhoon IP (GE Healthcare Life Sciences). Phosphorylated primer A-2 was used as a loading control. Activity was quantified as total radioactive incorporated per lane normalized to the loading control. To compare activity between 3xFLAG- and 3xFLAG-HaloTag containing telomerase, specific activity was calculated by dividing the activity measured by telomerase assays by the TR levels determined by northern blot. Processivity was quantified as the intensity of products longer than 6 repeats divided by the total signal per lane.
Patrick E.M., Slivka J., Payne B., Comstock M.J, & Schmidt J.C. (2020). Observation of processive telomerase catalysis using high-resolution optical tweezers. Nature chemical biology, 16(7), 801-809.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!