![(A) Schematic representation of the distribution of domains in the DENV NS5 protein. The FL NS5 protein has three major functional domains. The N-terminal MTase spans residues 1 to 296. The NLS has been divided into αNLS (spanning residues 320 to 368) and α/βNLS (residues 369 to 405). The arrow indicates the αNLS region thought to interact with the NS3 viral helicase. The six conserved amino acid sequence motifs within the RdRp domain (residues 273 to 900) are colored as in Fig. Fig.2C,2C, and “P” denotes the priming loop site. Also shown are the various truncated constructs of NS5 (see the text for details). (B) Schematic representation of the polymerase assay in the <t>SPA</t> bead format. The newly transcribed RNAs with the incorporation of <t>radioactive</t> <t>GTP</t> were captured on streptavidin-coupled beads and detected by counting as described in Materials and Methods. (C) Steady-state kinetics of RNA synthesis by DENV 3 RdRp, using a poly(C) primer and oligo(G) template. The reaction mixture contained 1 μM enzyme, 0.25 μg poly(C), 0.0125 μg oligo(G), 4 μM total GTP, and 0.5 μCi of [3H]GTP, as described in Materials and Methods. (D) Kinetic analysis of 3′dGTP inhibition. The IC50 of 3′dGTP for DENV 3 RdRp was determined using at least two independent experiments. Assay conditions are described in Materials and Methods. (E) Limited proteolysis of the DENV 2 FL NS5 protein (left) and of the DENV 3 RdRp domain (right). Lanes 1 and 4, molecular mass markers; lanes 2 and 3, FL undigested NS5 and trypsin-digested NS5, respectively; lanes 5 and 6, undigested DENV 3 RdRp and trypsin-digested DENV 3 RdRp, respectively. The trypsin/enzyme ratio for FL NS5 was 1:100, and the reaction was carried out at room temperature for 2 h (lane 3). Similarly treated DENV 3 RdRp was completely digested (data not shown). Using a lower concentration of trypsin (1:1,000 ratio of trypsin to RdRp) for ∼30 min at room temperature yielded several smaller fragments (lane 6). The N termini of digested protein fragments were sequenced (indicated by arrows), and this information was used to derive some of the constructs shown in panel A.](https://pub-med-central-thumbnail-images-cdn.bioz.com/pub_med_central_ids_ending_with_0186/pmc01900186/pmc01900186__zjv0090790770001.thumbnail.jpg)
![Poly(A) <t>SPA</t> assay principle. The PAP enzyme generates a polyadenylated RNA molecule (step i). Biotinylated oligo-d(T)20 oligonucleotide hybridises to the poly(A) tail and is reverse transcribed by Expand-RT in the presence of dNTPs and [3H]TTP (step ii). The radioactive, biotinylated RT product binds to <t>streptavidin</t> coated SPA beads (step iii) and can be directly measured by scintillation counting (step iv).](https://pub-med-central-thumbnail-images-cdn.bioz.com/pub_med_central_ids_ending_with_5778/pmc00055778/pmc00055778__gke40801.thumbnail.jpg)
![(A) SRF competes with colchicine for binding to microtubules as was determined using competition-binding scintillation proximity assay <t>(SPA).</t> In comparison to control (no competitor), SRF decreased colchicine binding by 2.5-fold. Data are shown as means ± SEM. * P <0.05, ** P <0.01 versus control. (B) SRF <t>and</t> <t>vinblastine</t> have distinct binding sites on tubulin as SRF does not compete with vinblastine. Results shown are mean (± SD) of three independent experiments. Data are shown as means ± SEM. * P <0.05 versus control. (C–E) Cartoon representation of binding mode of colchicine (C) and SRF (D) to tubulin. The binding mode of drugs to tubulin was examined by docking studies using molecular docking program GOLD (Genetic Optimization for Ligand Docking, Cambridge Crystallographic Data Centre, UK) . The compounds (colchicine or SRF) were docked into the colchicine-binding pocket of tubulin using the crystal structure of the tubulin-colchicine: stathmin-like domain [PDB code: 1SA0]. H-bond interactions between the indazole ring of SRF and N349 and K352 of β-chain (green) as well as the T179 and V181 of α-chain (blue) are highlighted. Panel E shows overlay of SRF and colchicine highlighting the difference in binding patterns of these molecules.](https://pub-med-central-thumbnail-images-cdn.bioz.com/pub_med_central_ids_ending_with_2991/pmc04212991/pmc04212991__pone.0110955.g002.thumbnail.jpg)
assistant_photoSave product to folder
X
Create new folder
All (7)
Amersham Life Sciences
Amersham Pharmacia Biotech
Cytiva
Revvity
Supercritical Fluid Technologies
BDO
Biacore
About 61 products / 201 snippets / 3 images
 86 / 100 Bioz Stars | Scintillation proximity assay (SPA) Danaher / Amersham   | star_border | Visit Supplier | |||
Article Snippets ".. The utilization of a scintillation proximity assay (SPA) ( 20 ) by using lectin-coated neuropeptide γ receptor SPA beads (Amersham Pharmacia) for CRFR binding analysis .." (see more) | × Open an account star_border Product associated with above image that matches your search query Danaher / Amersham Article Snippet "" Figure Legend " " | |||||
80 / 100 Bioz Stars | Phosphodiesterase scintillation proximity (SPA) beads Amersham Pharmacia Biotech Ltd | star_border | Visit Supplier | |||
Article Snippets ".. tRNA and ATP were purchased from Boehringer-Mannheim, L-[methyl-3H]methionine and phosphodiesterase scintillation proximity (SPA) beads from Amersham Pharmacia Biotech and other reagents from Sigma." (see more) | × | |||||
×![]()
80 / 100 Bioz Stars | Scintillation proximity assay (SPA) beads Revvity | star_border | Visit Supplier | |||
Article Snippets ".. the Human Calcitonin Receptor The binding assay was performed using scintillation proximity assay (SPA) beads (RPNQ0001) from PerkinElmer and cell membranes prepared from a BHK tk'ts 13 cell .." (see more) | × Open an account star_border Product associated with above image that matches your search query Article Snippet "" Figure Legend " " | |||||
80 / 100 Bioz Stars | Scintillation Proximity Assay (SPA) Beads Revvity | star_border | Visit Supplier | |||
Article Snippets ".. C. The enzyme reaction was stopped by the addition of Scintillation Proximity Assay (SPA) Beads (Perkin Elmer, USA) containing 5 mM 3-isobutyl-1-methylxanthine (IBMX)." (see more) | × Open an account star_border Product associated with above image that matches your search query Article Snippet "" Figure Legend " " | |||||
80 / 100 Bioz Stars | SPA (Scintillation Proximity Assay) beads Amersham Life Sciences Inc | star_border | Visit Supplier | |||
Article Snippets ".. After incubation for 18-24 hours at 37° C., the reaction is stopped adding 100 μl of Amersham SPA (Scintillation Proximity Assay) beads coupled to streptavidin, and the radioactivity .." (see more) | × | |||||
91 / 100 Bioz Stars | Scintillation proximity assay (SPA) beads Danaher / Cytiva | star_border | Visit Supplier | |||
Article Snippets "To measure the 50% inhibitory concentration (IC 50 ) of the 3′dGTP inhibitor, the enzyme was preincubated with template and inhibitor for 1 h before the reaction was started by the addition of a solution containing radioactive GTP for 1 h. The reaction was stopped .." (see more) | × Open an account
| |||||
80 / 100 Bioz Stars | Scintillation Proximity Assay (SPA) system Amersham Life Sciences Inc | star_border | Visit Supplier | |||
Article Snippets "A Scintillation Proximity Assay (SPA) system (Amersham International) is used to assess the incorporation of 32P radioactivity into CREB or CREBtide." (see more) | × | |||||
80 / 100 Bioz Stars | Scintilation proximity assay (SPA) beads Amersham Life Sciences Inc | star_border | Visit Supplier | |||
Article Snippets "After addition of 20 μA scintilation proximity assay (SPA) beads (100 μg/well, Amersham, RPNQ0001) components are gently mixed and incubated at RT .." (see more) | × | |||||
80 / 100 Bioz Stars | Scintillation proximity assay (SPA) kit Amersham Life Sciences Inc | star_border | Visit Supplier | |||
Article Snippets ".. with the Amersham kit) and the cyclic AMP content was determined using an Amersham scintillation proximity assay (SPA) kit, following the manufacturer's instructions." (see more) | × | |||||
80 / 100 Bioz Stars | Scintilation proximity assays (SPA) Biacore Biacore | star_border | Visit Supplier | |||
Article Snippets ".. assays include, but are not limited to, ELISA, fluorescence anisotropy and intensity, scintilation proximity assays (SPA) Biacore (Pearce et al., Biochemistry 38: 81-89 (1999)), DELFIA assays, and AlphaScreenTM (commercially .." (see more) | × | |||||
80 / 100 Bioz Stars | Scintillation proximity assay (SPA) technology Amersham Life Sciences Inc | star_border | Visit Supplier | |||
Article Snippets ".. tested for their ability to inhibit Src kinase using the scintillation proximity assay (SPA) technology as developed by Amersham." (see more) | × | |||||
80 / 100 Bioz Stars | Anti Scintillation proximity assay (SPA) beads Amersham Pharmacia Biotech Ltd | star_border | Visit Supplier | |||
Article Snippets "Pharmacological Methods Materials: 3H-cortisone, 3H-cortisol and scintillation proximity assay (SPA) beads were purchased from Amersham Pharmacia Biotech, β-NADPH and β-NAD were from Sigma Chemical .." (see more) | × | |||||