Spa beads

Manufactured by PerkinElmer

SPA beads are a type of lab equipment used in various applications. They serve as a platform for performing scintillation proximity assays, which rely on the principle of energy transfer to detect biomolecular interactions. The beads are designed to efficiently capture and detect radioactive signals, enabling researchers to analyze and quantify specific analytes or targets within a sample.

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Lab products found in correlation

Radioactive isotopes, by PerkinElmer (5 mentions) 96 well screening plates, by PerkinElmer (4 mentions) Penicillin streptomycin solution, by Corning (2 mentions) 3h sam, by PerkinElmer (1 mentions) Oligonucleotide, by Integrated DNA Technologies (1 mentions) Dna oligonucleotide, by Integrated DNA Technologies (1 mentions) 96 well plate, by PerkinElmer (1 mentions)

Spelling variants of this product

Yttrium SPA beads (4 citations) Ysi) poly-L-lysine coated SPA beads (3 citations) PVT WGA SPA beads (3 citations) RNA Binding YSi SPA Beads (2 citations) Copper-chelated SPA beads (2 citations) Streptavidin-coated PVT SPA beads (2 citations) YSi scintillation proximity assay beads (SPA, (2 citations)

7 protocols using spa beads

High-throughput DNMT1 Inhibitor Screening

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A homogenous 384-well scintillation proximity assay (SPA) was developed and optimized for screening of DNMT-1 inhibitors. In this assay ³H-SAM (PerkinElmer) is used as a methyl-group donor in the methylation of a biotinylated 42mer hemi-methylated substrate comprising the following two oligonucleotides (IBA): 5′ (bio-LC)GAT CCG ACG ACG ACG ACG AXG ACG ACG ACG ACG ACG ACG ATC (X = 5 methylcytosine) and 5′GAT CGT CGT CGT CGT CGT CGT CGT CGT CGT CGT CGT CGG ATC. The methylated is captured with yttrium silicate (YSi) streptavidin coated SPA beads (PerkinElmer). The library containing the subsets: FDA approved drugs (Enzo Life Sciences), Naturstoffe (Enzo Life Sciences), Natural products (Cfm Tropitzsch), Flavonoid derivatives (TimTec) and a collection of additional compounds (Beiersdorf AG) was screened on two separate experimental days against recombinant human DNMT1 at the DKFZ/EMBL Chemical Biology Core Facility. Compounds/substances were diluted 1:25 in H₂O and dispensed into 384-well Optiplates. 800 nM ³H-SAM was added to 100 nM 42mer oligo and 30 nM DNMT1 to start the methylation reaction (180 min at room temperature). Subsequently, YSi beads were added with 372 mM NaCl to stop the methylation reaction and the plates were read on a MicroBeta LumiJet.

Falckenhayn C., Bienkowska A., Söhle J., Wegner K., Raddatz G., Kristof B., Kuck D., Siegner R., Kaufmann R., Korn J., Baumann S., Lange D., Schepky A., Völzke H., Kaderali L., Winnefeld M., Lyko F, & Grönniger E. (2024). Identification of dihydromyricetin as a natural DNA methylation inhibitor with rejuvenating activity in human skin. Frontiers in Aging, 4, 1258184.

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Antimicrobial Compound Screening Protocol

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Oligonucleotides were from Integrated DNA Technologies (Coralville, IA). All other chemicals were obtained from either Sigma Aldrich (St. Louis, MO) or Fisher Scientific (Pittsburgh, PA). DNA sequencing was performed by Functional Bioscience (Madison, WI). Radioactive isotopes, SPA beads, and 96-well screening plates were from PerkinElmer (Waltham, MA). Escherichia coli tolC mutant, P. aeruginosa PAO200 (efflux pump mutant), and P. aeruginosa hypersensitive strain (ATCC 35151) were a kind gift from Urs Ochsner (Crestone Pharma, Boulder, CO). All other bacteria were from the American Type Culture Collection (ATCC) (Manassas, VA). Penicillin-Streptomycin Solution was from Mediatech, Inc. (Centerville, IA). The synthetic compound library was from TimTec LLC (Newark, DE), and the natural compound library was from MicroSource Discovery Systems (Gaylordsville, CT).

Hu Y., Guerrero E., Keniry M., Manrrique J, & Bullard J.M. (2015). Identification of Chemical Compounds That Inhibit the Function of Glutamyl-tRNA Synthetase from Pseudomonas aeruginosa. Journal of biomolecular screening, 20(9), 1160-1170.

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High-Throughput Antimicrobial Screening

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All chemicals were obtained from Fisher Scientific (Pittsburg, PA). Radioactive isotopes, SPA beads and 96-well screening plates were from PerkinElmer (Waltham, MA). E. coli tolC mutant, P. aeruginosa PAO200 (efflux pump mutant) and P. aeruginosa hypersensitive (ATCC^® 35151^™) strains were a kind gift from Urs Ochsner (Crestone Pharma, Boulder, CO). All other bacteria were from American Type Culture Collection (ATCC) (Manassas, VA). The synthetic compound library was from TimTec LLC (Newark, DE) and the natural product library was from MicroSourceDiscovery Systems, Inc. (Gaylordsville, CT). Compounds were supplied as 10 mM stocks dissolved in dimethyl sulfoxide (DMSO), stored at −20 °C and thawed immediately before analysis. The compounds have an average purity of 95%, and the minimum purity is at least 90%.

Robles S., Hu Y., Resto T., Dean F, & Bullard J.M. (2017). Identification and Characterization of a Chemical Compound that Inhibits Methionyl-tRNA Synthetase from Pseudomonas aeruginosa. Current drug discovery technologies, 14(3), 156-168.

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High-throughput screening of synthetic compounds

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All chemicals were obtained from either Sigma Aldrich (St. Louis, MO) or Fisher Scientific (Pittsburgh, PA). Radioactive isotopes, SPA beads, and 96-well screening plates were from PerkinElmer (Waltham, MA). Escherichia coli tolC mutant, P. aeruginosa PAO200 (efflux pump mutant), and P. aeruginosa hypersensitive (ATCC 35151) strains were a kind gift from Urs Ochsner (Crestone Pharma, Boulder, CO). All other bacteria were from the American Type Culture Collection (ATCC) (Manassas, VA). Penicillin-streptomycin solution was from Mediatech, Inc. (Centerville, IA). The synthetic Anti-Infectives Library was from TimTec LLC (Newark, DE) and included 890 low-molecular-weight, druglike molecules with scaffolds found in antiseptic agents with antibacterial, antifungoid, and antimicrobial activities. Compounds were supplied as 10 mM stocks dissolved in DMSO, stored at −20 °C, and thawed immediately before analysis. The compounds have an average purity of 95%, and the minimum purity is at least 90%.

Palmer S.O., Hu Y., Keniry M, & Bullard J.M. (2016). Identification of Chemical Compounds That Inhibit Protein Synthesis in Pseudomonas aeruginosa. SLAS discovery, 22(6), 775-782.

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Chemical Reagents and Compound Library

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All chemicals were obtained from Fisher Scientific (Pittsburgh, PA). DNA oligonucleotides were from Integrated DNA Technologies (Coralville, IA). DNA sequencing was performed by Functional Bioscience (Madison, WI). Radioactive isotopes, SPA beads, and 96-well screening plates were from PerkinElmer (Waltham, MA). The Soluble Diversity chemical compound library was from ChemDiv, Inc. (San Diego, CA). Compounds stocks were dissolved in DMSO to a concentration of 10 mM, stored at −20 °C, and thawed immediately before analysis. The compounds had a minimum purity of at least 90%.

Hughes C.A., Gorabi V., Escamilla Y., Dean F.B, & Bullard J.M. (2020). Two Forms of Tyrosyl-tRNA Synthetase from Pseudomonas aeruginosa: Characterization and Discovery of Inhibitory Compounds. SLAS discovery : advancing life sciences R & D, 25(9), 1072-1086.

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High-Throughput Compound Screening

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All chemical compounds used in this study were obtained from Fisher Scientific (Pittsburg, PA). Radioactive isotopes, SPA beads, and 96-well plates were obtained from PerkinElmer (Waltham, MA). The synthetic compound library was from TimTec LLC (Newark, DE), and the natural product library was from MicroSourceDiscovery Systems, Inc. (Gaylordsville, CT). The chemical compounds were obtained as 10 mM stock solutions dissolved in DMSO and stored at −20 °C. These compounds were thawed immediately before screening. The compounds have an average purity of 95%, with the minimum purity at least 90%.

Balboa S., Hu Y., Dean F.B, & Bullard J.M. (2019). Lysyl-tRNA Synthetase from Pseudomonas aeruginosa: Characterization and Identification of Inhibitory Compounds. SLAS discovery : advancing life sciences R & D, 25(1), 57-69.

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Quantifying SAM-binding Methyltransferase Activity

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Biotinylated DNV3MTase (580 nM) was mixed with the polyvinyltoluene (PVT) scintillation proximity assay (SPA) beads (1.5 mg/mL, PerkinElmer) and 20 μM of SAM, sinefungin (SIN), or each compound in the SAM binding buffer (20 mM Tris pH 7.5, 50 mM NaCl, 10 mM KCl, 2 mM MgCl₂, 2 mM MnCl₂, 0.05% CHAPS) in a white 96-well clear-bottom plate. The samples were mixed by gentle rocking for 20 min at 23°C, followed by the addition of 1.65 μCi of ³H-SAM (425 nM) to a final sample volume of 50 μL. After mixing for another 15 min at 23°C, samples were then centrifuged for 2 min at 500g and analyzed on a Microbeta^{2 (link)} 2450 plate counter (PerkinElmer) using the default ³H-Scintillation Proximity Assay protocol within the manufactory software.

Vernekar S.K., Qiu L., Zhang J., Kankanala J., Li H., Geraghty R.J, & Wang Z. (2015). 5′-Silylated 3′-1,2,3-triazolyl Thymidine Analogues as Inhibitors of West Nile Virus and Dengue Virus. Journal of medicinal chemistry, 58(9), 4016-4028.

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