Wallac 1450 microbeta counter

Manufactured by PerkinElmer

Sourced in United Kingdom, Italy

The Wallac 1450 MicroBeta counter is a laboratory instrument designed for liquid scintillation counting. It is used to measure the radioactivity of samples by detecting the light signals produced when radioactive particles interact with a scintillation cocktail.

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Lab products found in correlation

Whatman, by Cytiva (4 mentions) Prism 7, by GraphPad (1 mentions) Gf c filters, by Brandel Inc (1 mentions) Glass microfiber gf c filters, by Cytiva (1 mentions) Ecolume liquid scintillation cocktail, by MP Biomedicals (1 mentions) 24 well harvester, by Brandel Inc (1 mentions) Prism 6, by GraphPad (1 mentions)

Spelling variants of this product

MicroBeta counter (40 citations) 1450 Microbeta (18 citations) Wallac 1450 MicroBeta (13 citations) Wallac Microbeta (9 citations) Wallac MicroBeta counter (8 citations) Wallac 1450 MicroBeta TriLux Liquid Scintillation Counter (6 citations) Wallac 1450 Microbeta liquid scintillation counter (5 citations) Wallac 1450 Microbeta scintillation counter (3 citations) P-E 1450 Microbeta Wallac Trilux scintillation counter (3 citations) Wallac 1450 Microbeta Trilux β-counter (2 citations)

6 protocols using wallac 1450 microbeta counter

Hippocampal 5HT1A Receptor Binding Assay

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Hippocampal membrane homogenates (100 μg protein/well) were incubated in 50 mM pH 7.4 Tris-HCl buffer with eight concentrations of the 5HT1A receptor ligand [³H] 8-OH-DPAT ranging in concentration from 0.16 nM to 20 nM. Non-specific binding was determined using 10 μM WAY-100635. The assay was incubated for 60 min at room temperature before filtration through a Whatman GF/C filters using a MLR-24 Brandel harvester. Bound radioactivity was then determined by scintillation counting with a Wallac 1450 Microbeta counter (Perkin Elmer). Each condition was performed in triplicate and independently replicated three times. Data were analyzed using GraphPad Prism 7.0 (GraphPad; La Jolla, CA).

Senese N., Oginsky M., Neubig R.R., Ferrario C., Jutkiewicz E.M, & Traynor J.R. (2018). Role of hippocampal 5-HT1A receptors in the antidepressant-like phenotype of mice expressing RGS-insensitive Gαi2 protein. Neuropharmacology, 141, 296-304.

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Adipocyte Glucose Uptake Assay

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hASC were seeded in Cytostar T 96-well plates at 15,000 cells/well density and fully differentiated in adipocyte medium (AM; Zen-Bio Inc.) with 0.05 mM IBMX, 0.1 μM dexamethasone, 10 nM insulin, and 1 μM pioglitazone (7 days); and thereafter kept in AM (changed every other day). For glucose uptake, adipocytes were starved for 3 h in DMEM/0.1% BSA, stimulated with FGF-21 for 24 h, and washed twice with KRP buffer (15 mM HEPES, pH 7.4, 118 mM NaCl, 4.8 mM KCl, 1.2 mM MgSO₄, 1.3 mM CaCl₂, 1.2 mM KH₂PO₄, 0.1% BSA), and 100 μL of KRP buffer containing 2-deoxy-d-14Cglucose (2-DOG) (0.1 μCi, 100 μM) was added to each well. Control wells contained 100 μL KRP buffer with 2-DOG (0.1 μCi, 10 mM) to monitor for nonspecificity. The uptake reaction was carried out for 1 h at 37°C, and measured using a Wallac 1450 MicroBeta counter (Perkin Elmer).

Bartesaghi S., Wallenius K., Hovdal D., Liljeblad M., Wallin S., Dekker N., Barlind L., Davies N., Seeliger F., Winzell M.S., Patel S., Theisen M., Brito L., Bergenhem N., Andersson S, & Peng X.R. (2022). Subcutaneous delivery of FGF21 mRNA therapy reverses obesity, insulin resistance, and hepatic steatosis in diet-induced obese mice. Molecular Therapy. Nucleic Acids, 28, 500-513.

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Cell Membrane Binding Assay

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Cell membranes (as prepared above, 10 μg/well) were incubated in the following mixture for 90 min at 25 °C: assay buffer (50 mM Tris-HCl, pH 7.4, 100 mM NaCl, 1 mM EDTA, 5 mM MgCl₂, 10 μM GTPγS), various concentrations of orthosteric and allosteric ligand, and 0.35-0.45 nM [³H]DPN. Nonspecific binding was determined in the presence of 10 μM naloxone. Reactions were terminated by rapid filtration through glass microfiber GF/C filters (Whatman) using a Brandell harvester and washed three times using cold 50 mM Tris-HCl buffer. Filters were dried in a 50 °C oven, and radioactivity was measured by liquid scintillation counting with EcoLume liquid scintillation cocktail (MP Biomedicals) in a Wallac 1450 MicroBeta counter (PerkinElmer).

Burford N.T., Livingston K.E., Canals M., Ryan M.R., Budenholzer L.M., Han Y., Shang Y., Herbst J.J., O'Connell J., Banks M., Zhang L., Filizola M., Bassoni D.L., Wehrman T.S., Christopoulos A., Traynor J.R., Gerritz S.W, & Alt A. (2015). Discovery, Synthesis, and Molecular Pharmacology of Selective Positive Allosteric Modulators of the δ-Opioid Receptor. Journal of medicinal chemistry, 58(10), 4220-4229.

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Opioid Receptor Binding Assay

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For competition binding experiments in µ-OR- rHDL, a mixture of µ-OR-rHDL and ³H-diprenorphine (³H-DPN) was incubated with varying concentrations of agonist in a binding buffer comprised of 25 mM HEPES pH 7.4, 100 mM NaCl, and 0.1% BSA in the presence or absence of 3 µM Nb39. For assays performed using cell membranes, conditions listed were kept the same except for exclusion of BSA and inclusion of 10 µg protein per well. Binding reactions were incubated for 2 h at 25°C. Free radioligand was separated from bound radioligand by rapid filtration onto a Whatman GF/C filter pretreated with 0.1% polyethylenimine using a 24-well harvester (Brandel). Nonspecific binding was measured in the presence of 10 µM naloxone, an opioid antagonist. Radioligand activity was measured by liquid scintillation counting using a Wallac 1450 MicroBeta counter (Perkin Elmer). A minimum of three independent experiments, each in duplicate, were performed and the values were pooled to generate the mean curves displayed in the figures. Competition binding data were fit to a one-site model using GraphPad Prism 6.0. Data are presented as means with 95% confidence limits in parentheses. No statistical methods were used to predetermine sample size.

Livingston K.E., Mahoney J.P., Manglik A., Sunahara R.K, & Traynor J.R. (2018). Measuring ligand efficacy at the mu-opioid receptor using a conformational biosensor. eLife, 7, e32499.

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Monitoring Gα subunit activation in vitro

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Incorporation of the slowly-hydrolyzed GTP analog [³⁵S]GTPγS into activated Gα subunits was monitored in vitro (Traynor and Nahorski 1995 (link)). Brain homogenates prepared as above (10 µg protein) were incubated for 2 h at 25°C in [³⁵S]GTPγS binding buffer (50 mM Tris, 5 mM MgCl₂, 100 mM NaCl and 1 mM EDTA, pH 7.4, with 2 mM dithiothreitol, 100 µM GDP and 0.4 U/mL adenosine deaminase) with 0.1 nM [³⁵S]GTPγS and a maximal concentration (10 µM) of either DAMGO or morphine. Non-specific binding was evaluated in the presence of 10 µM unlabeled GTPγS. Reactions were stopped by rapid filtration through GF/C filtermats (Whatman, Kent, UK) using a Brandel MLR-24 harvester (Brandel, Gaithersburg, MD), and bound radioactivity was determined by liquid scintillation counting using a Wallac 1450 MicroBeta counter (PerkinElmer, Waltham, MA).

Lamberts J.T., Rosenthal L.D., Jutkiewicz E, & Traynor J.R. (2017). Role of the guanine nucleotide binding protein, Gαo, in the development of morphine tolerance and dependence. Psychopharmacology, 235(1), 71-82.

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Evaluating ROS Production by Luminol Assay

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ROS production by HaCaT cells exposed to GBNs was also evaluated by the luminol assay. Preliminarily, the potential interference of FLG and GO with the assay was evaluated in an acellular system by incubating each GBN (0.4-100 μg mL -1 ) with luminol for 15 minutes at 37 °C. Subsequently, the cells were seeded overnight at a density of 1 × 10 4 cells per well in 96-well white plates and exposed to GBNs (0.4-100 μg mL -1 ) for 24 h before adding luminol (1 μM) to each well. Chemiluminescence was recorded 15 minutes after luminol addition, by using a multiwell luminometer (Wallac 1450 Microbeta counter, PerkinElmer, Milan, Italy). The results are expressed as % of ROS production as compared to negative control (cells not exposed to GBNs).

Pelin M., Fusco L., Martín C., Sosa S., Frontiñán-Rubio J., González-Domínguez J.M., Durán-Prado M., Vázquez E., Prato M, & Tubaro A. (2018). Graphene and graphene oxide induce ROS production in human HaCaT skin keratinocytes: the role of xanthine oxidase and NADH dehydrogenase. Nanoscale, 10(25).

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