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Amersham hyperfilm mp

Manufactured by GE Healthcare
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Amersham Hyperfilm MP is a high-performance X-ray film designed for a wide range of laboratory applications. It provides consistent and reliable results, making it a versatile choice for researchers and scientists.

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Lab products found in correlation

Prime it 2 random primer labeling kit, by Agilent Technologies (5 mentions) Hybond n membrane, by GE Healthcare (5 mentions) Bas ip ms storage phosphor screen, by GE Healthcare (3 mentions) 3 3 5 5 tetramethylbenzidine tmb peroxidase substrate, by LGC (3 mentions) Can get signal solutions 1 and 2, by Toyobo (3 mentions) Hrp conjugated secondary antibody, by Agilent Technologies (2 mentions) Clarity western ecl, by Bio-Rad (2 mentions) Typhoon fla 7000 phosphorimager, by GE Healthcare (2 mentions) Chemidoc xrs system, by Bio-Rad (2 mentions) Random primed dna labelling kit, by Roche (2 mentions) α 32p dctp, by PerkinElmer (2 mentions) Hrp conjugated goat anti mouse igg1 antibody, by Fortis Life Sciences (2 mentions) Bradford protein assay reagent, by Nacalai Tesque (2 mentions) Pentobarbital sodium salt, by Nacalai Tesque (2 mentions) 3mm chromatography paper, by GE Healthcare (2 mentions) Pk11195, by Merck Group (2 mentions) Maxiscript kit, by Thermo Fisher Scientific (2 mentions) Fla 7000 phosphoimager, by Fujifilm (2 mentions) Typhoon phosphorimager, by GE Healthcare (2 mentions) Western blotting luminol reagent, by Santa Cruz Biotechnology (2 mentions)

Spelling variants of this product

Hyperfilm (131 citations) Amersham Hyperfilm (89 citations) Hyperfilm MP (29 citations) Amersham Hyperfilm MP film (3 citations) Amersham Hyperfilm ECL and MP Autoradiography Films (2 citations) Amersham Hyperfilm MP autoradiography film (2 citations)

44 protocols using amersham hyperfilm mp

1

Radioactive Fatty Acid Labeling

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Cells were seeded in tissue culture dishes as described above. For labelling, the cells were starved using IM medium (Glasgow minimal essential medium buffered with 10 mM Hepes, pH 7.4). After 1 h, the medium was replaced by IM with 3H-palmitate at 200 µCi/mL (American Radiolabeled Chemicals, US) for 2 h at 37 °C. Cell lysis, immunoprecipitation and SDS-PAGE were performed as above. The gels were fixed for 30 min with 10% acetic acid, 25% isopropanol in water and the signal was amplified for 30 min with NAMP100 (GE Healthcare, US). The gels were then dried and applied to an Amersham Hyperfilm MP (GE Healthcare, US). The radioactivity was visualized and quantify using a Typhoon TRIO (GE Healthcare, US).
Sandoz P.A., Denhardt-Eriksson R.A., Abrami L., Abriata L.A., Spreemann G., Maclachlan C., Ho S., Kunz B., Hess K., Knott G., S. Mesquita F., Hatzimanikatis V, & van der Goot F.G. (2023). Dynamics of CLIMP-63 S-acylation control ER morphology. Nature Communications, 14, 264.
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2

Enzyme-linked Immunoassay for Diisocyanate Exposure

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H2O2, the diisocyanates, 4,4′-diphenylmethane diisocyanate (MDI), 1,6 hexamethylene diisocyanate (HDI), and an 80:20 ratio mixture of 2,4:2,6 toluene diisocyanate (TDI), albumins (from human, bovine and mouse), tropomyosin, keyhole-limpet hemocyanin (KLH), Tween 20, tetramethylbenzidine (TMB) liquid substrate solution, complete and incomplete Freund’s adjuvant were from Sigma-Aldrich (St. Louis, MO). Biotin-labeled mouse monoclonal anti-rabbit IgG (6C1A8) with no cross-reactivity to IgG from other species was from ProSci Inc. (Poway, CA). Streptavidin-HRP and peroxidase conjugated anti-rabbit IgG were from BD Biosciences (San Jose, CA). Biotinylated goat anti-human albumin was from Bethyl (Montgomery, TX) and protein-A purified anti-IL-33 polyclonal rabbit IgG was from Enzo Life Sciences (Farmingdale, NY). Cyanogen bromide activated sepharose 4B was from Amersham Biosciences (Uppsala, Sweden) and protein A affinity columns were from BioRad (Hercules, CA). Biotin blocking solution was from Life Technologies (Eugene, OR). Diaminobenzidine (DAB) substrate, hematoxylin, and histomount were from a HistoMouse-MAX staining Kit (Invitrogen, Frederick, MD). Protein A sepharose 4B (InVitrogen). Normal rabbit and goat serum were from ThermoFisher Scientific (Waltham, MA). Amersham Hyperfilm MP from GE Healthcare Bio-Sciences (Pittsburgh, PA) was used in developing dot blots.
Wisnewski A.V, & Liu J. (2015). Immunochemical detection of the occupational allergen, methylene diphenyl diisocyanate (MDI), in situ. Journal of immunological methods, 429, 60-65.
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3

Comprehensive Western Blot Analysis Protocol

Cited 2 times
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Western blotting was carried out as described previously [11 (link), 12 (link)]. Whole cell extracts (50 µg/lane) were dissolved on 7.5% acrylamide gels and transferred onto Polyvinyl difluoride (PVDF) membranes (Immobilon-P, 0.45 µm, Merck Millipore). Primary antibodies (Table 1) were diluted 1:1000 (except anti-ARNT 1:2000) and applied overnight. Determination of Lamin A/C was done for normalisation and the appropriate antibody was applied for 1 h at room temperature. Afterwards PVDF membranes were incubated with HRP-conjugated secondary antibodies (1:5000, DAKO) for 1 h at room temperature. Chemoluminescence development was achieved using the ECL reagent (Clarity™ Western ECL, Bio-Rad). Subsequently membranes were exposed to X-ray films (Amersham Hyperfilm MP, GE Healthcare) and signals were quantified using the AIDA Image Analyzer (Version 4.27, Raytest).
Mandl M, & Depping R. (2017). ARNT is a potential direct HIF-1 target gene in human Hep3B hepatocellular carcinoma cells. Cancer Cell International, 17, 77.
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4

Quantifying Serotonin Transporter Binding

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Serotonin transporter (SERT) binding was determined on brain sections, cut as above, using (3H)-paroxetine (Perkin Elmer, UK) as previously described [61 (link)]. Slides were then exposed to (3H)-sensitive film (Amersham Hyperfilm MP, GE Healthcare, UK) at −80°C for 6 weeks. Analysis of autoradiographs was performed by measuring the signal over the area of interest with densitometry software (MCID Basic 7.0, Imaging Research, Inc.). The background was subtracted.
Issler O., Carter R.N., Paul E.D., Kelly P.A., Olverman H.J., Neufeld-Cohen A., Kuperman Y., Lowry C.A., Seckl J.R., Chen A, & Jamieson P.M. (2014). Increased anxiety in corticotropin-releasing factor type 2 receptor-null mice requires recent acute stress exposure and is associated with dysregulated serotonergic activity in limbic brain areas. Biology of Mood & Anxiety Disorders, 4, 1.
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5

Imaging Techniques for Protein Analysis

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Ethidium bromide-stained gels and western blots were imaged with a BioRad ChemiDoc XRS + system.
Gels containing radioactive samples were dried onto chromatography paper (GE Healthcare, 3030-861) and imaged with a Typhoon FLA 7000 phosphorimager (GE Healthcare) for quantitative analyses. Dried gels were also exposed to Amersham Hyperfilm MP (GE Healthcare, 28906843) and scanned for data presentation.
Brüning J.G, & Marians K.J. (2021). Bypass of complex co-directional replication-transcription collisions by replisome skipping. Nucleic Acids Research, 49(17), 9870-9885.
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6

Quantitative Immunoblot Analysis

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Protein expression on immunoblots was quantified on 8-bit gray-scale-transformed.tiff images of either scanned Amersham Hyperfilm MP (GE Healthcare, 28906838) or.tiff images obtained by the Bio-Rad ChemiDoc imaging system. Fiji ImageJ was used to select a region of interest for the respective proteins for densitometric analysis. Protein expression for each protein was normalized to the loading control of the respective sample.
Apriamashvili G., Vredevoogd D.W., Krijgsman O., Bleijerveld O.B., Ligtenberg M.A., de Bruijn B., Boshuizen J., Traets J.J., D’Empaire Altimari D., van Vliet A., Lin C.P., Visser N.L., Londino J.D., Sanchez-Hodge R., Oswalt L.E., Altinok S., Schisler J.C., Altelaar M, & Peeper D.S. (2022). Ubiquitin ligase STUB1 destabilizes IFNγ-receptor complex to suppress tumor IFNγ signaling. Nature Communications, 13, 1923.
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7

Northern Blot Assay for RNA Analysis

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Northern blot experiments were essentially performed as described (Vasiljeva & Buratowski, 2006 (link)). RNA was prepared as described in Kilchert et al (2015) (link). 8 μg of RNA were resolved on a 1.2% agarose gel containing 6.7% formaldehyde in MOPS buffer. After capillary transfer in 10× SSC onto a Hybond N+ membrane (GE Healthcare), RNA was UV-crosslinked and stained with methylene blue to visualise ribosomal RNAs. Gene-specific probes were generated by random priming in the presence of ATP [α32P] using the Prime-It II Random Primer Labeling Kit (300385; Agilent) using PCR-generated DNA templates produced from genomic DNA isolated from a wild type S. pombe strain (YP71) with the oligonucleotides listed in Table S11. Probes were added to the membrane and hybridised at 42°C overnight. After repeated washes in 2× SSC, 0.1% SDS, blots were exposed to Amersham Hyperfilm MP (28-9068-44; GE Healthcare).
Birot A., Kus K., Priest E., Al Alwash A., Castello A., Mohammed S., Vasiljeva L, & Kilchert C. (2021). RNA-binding protein Mub1 and the nuclear RNA exosome act to fine-tune environmental stress response. Life Science Alliance, 5(2), e202101111.
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8

IgE Detection in Patient Sera

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After separation by electrophoresis using a 15% polyacrylamide gel, proteins was blotted to a 0.45 µm nitrocellulose membrane (GE Healthcare, Uppsala, Sweden). Blotting was performed at 150 mA for 45 min. at 4 °C. Patients sera were diluted in TBS-T containing 1% BSA and 100 µg HRP as for ELISA. The secondary antibody, HRP conjugated goat anti-human IgE (KPL, Gaithersburg, MD, USA) was diluted 1:10000 in TBS-T + 1% BSA and incubated for 2 h at room temperature on the membrane. Blots were developed using the Clarity ECL Western Blot Substrate (Bio-Rad Laboratories, Inc.) following the manufacturers instruction. The membrane was placed in a light protected cassette (Hypercassette, Amersham, GE Healthcare, Japan) and exposed to an X-ray film (Amersham Hyperfilm™ MP; GE Healthcare, Japan) in the dark. Depending on the signal, films were exposed between 0.5 and 10 min and developed using a CP1000 AGFA film processor (Superior Radiographics, LTD., Madison, WI, USA).
Dubiela P., Kabasser S., Smargiasso N., Geiselhart S., Bublin M., Hafner C., Mazzucchelli G, & Hoffmann-Sommergruber K. (2018). Jug r 6 is the allergenic vicilin present in walnut responsible for IgE cross-reactivities to other tree nuts and seeds. Scientific Reports, 8, 11366.
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9

Quantitative Western Blot Analysis of hiPSC-CMs

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Western blot analysis was performed on total crude protein lysates from cultured hiPSC-CMs in the different conditions. Same amount of proteins (20 μg/lane) of single samples or pooled samples (n = 5–6) were separated on a 10% SDS-polyacrylamide (29:1) mini-gels (Bio-Rad) and transferred by wet-electroblotting to nitrocellulose membranes. Membranes were stained with the primary antibodies directed against the M-motif of cMyBP-C (polyclonal, 1:10,000, custom made) and α-actinin (monoclonal, 1:10,000, Sigma). Peroxidase-conjugated secondary antibodies against mouse (1:20,000, Dianova) or against rabbit (1:20,000, Sigma) were used. Proteins were visualized using Amersham ECL Prime Western Blotting Detection Reagent (GE Healthcare), and the signals were detected on Amersham Hyperfilm MP (GE Healthcare). Signals were quantified with GeneTools image analyzing software (Syngene).
Prondzynski M., Krämer E., Laufer S.D., Shibamiya A., Pless O., Flenner F., Müller O.J., Münch J., Redwood C., Hansen A., Patten M., Eschenhagen T., Mearini G, & Carrier L. (2017). Evaluation of MYBPC3 trans-Splicing and Gene Replacement as Therapeutic Options in Human iPSC-Derived Cardiomyocytes. Molecular Therapy. Nucleic Acids, 7, 475-486.
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10

RNA Analysis by Northern Blotting

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RNA extraction, gel electrophoresis, blotting, and detection with a radiolabeled probe were performed as described previously (29 (link)) but with the following adjustments. Ten micrograms of total RNA for each cell line was loaded onto the gel. The probes were labeled with the DecaLabel DNA labeling kit (Thermo Fisher Scientific), and the membrane was exposed to Amersham Hyperfilm MP (GE Healthcare) for 8 days (in the case of the SERINC5-specific probe) or for 8 h (glyceraldehyde-3-phosphate dehydrogenase [GAPDH]-specific probe) at −80°C. The SERINC5-specific probe of 870 bp was prepared by digestion of a plasmid bearing the SERINC5 cDNA with XbaI and NotI. For the loading control, a GAPDH plasmid (a gift from K. Habers, Heinrich-Pette-Institut, Hamburg, Germany) was digested by EcoRI, generating a 1.3-kbp GAPDH-specific probe.
Passos V., Zillinger T., Casartelli N., Wachs A.S., Xu S., Malassa A., Steppich K., Schilling H., Franz S., Todt D., Steinmann E., Sutter K., Dittmer U., Bohne J., Schwartz O., Barchet W, & Goffinet C. (2019). Characterization of Endogenous SERINC5 Protein as Anti-HIV-1 Factor. Journal of Virology, 93(24), e01221-19.
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