Counterflow assay protocol
WebSep 26, 2010 · Counterflow assay. All counterflow assays were carried out at 25 °C. Concentrated proteoliposomes (2 μl) were diluted into 100 μl assay buffer containing 1 μCi [3 H]-l-fucose (specific ... WebDec 3, 2024 · Establishment and Validation of Competitive Counterflow as a Method To Detect Substrates of the Organic Anion Transporting Polypeptide 2B1 ... It was the aim of this study to test whether CCF can be used to identify substrates of OATP2B1. A protocol for CCF ... Results of the CCF were complemented with cytotoxicity assays or …
Counterflow assay protocol
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WebAfter optimizing the protocol to be applied for OATP2B1 (for details refer to [1]), we screened a library of molecules for their influence on the cellular amount of estrone 3 … WebAug 28, 2024 · Using our counterflow assay, we confirmed the selective inhibition of PfHT1 by C3361. The IC 50 values of C3361 are 33.1 ± 2.0 μM for PfHT1, and approximately 1.3 mM and 0.7 mM for GLUT1 and GLUT3, respectively ... The rest of the protocol is identical to that described above. To determine IC 50 of the inhibitors, ...
WebIncubate cells in the diluted antibody in 1% BSA in PBST in a humidified chamber for 1 h at room temperature or overnight at 4°C. Decant the solution and wash the cells three times in PBS, 5 min each wash. Incubate cells with the secondary antibody in 1% BSA for 1 h at room temperature in the dark. Decant the secondary antibody solution and ... WebOct 31, 2024 · The competitive counterflow (CCF) assay does not require radiolabeled substrates or LC-MS/MS methods and is a more cost-effective approach to identifying …
WebSep 8, 2024 · However, the competitive counterflow (CCF) assay originally described for OCT2 and, more recently, for OCT1, OATP2B1, and OATP1A2 does not require radiolabeled substrates or LC-MS/MS … WebThe competitive counterflow assay measures the uptake of a test compound by measuring the concentration of the fluorescent indicator substrate 8-acetoxypyrene-1,3,6-trisulfonate ... (Protocol #2015A00000101-R2; approved 9/20/2024) and conducted in accordance with the NIH’s Guide for the Care and Use of Laboratory Animals.
WebThe organic anion transporting polypeptide (OATP) 2B1 is ubiquitously expressed and known to facilitate cellular entry. It is widely accepted that transport proteins play a pivotal role in pharmacokinetics. Consequently, testing for interaction with drug transporters became an important part in the assessment of new molecular entities in order to predict …
WebFeb 9, 2024 · Introducing the Gibco CTS Rotea Counterflow Centrifugation System, a closed cell processing system for cell therapy manufacturing. The Rotea system offers exceptional cell recovery, flexible input and output volume capability, and … grand banks power \u0026 motoryacht adWebCounterflow Centrifugation System. The bottom left corner of the log in page lists the software/firmware version number and the CTS Rotea Counterflow Centrifugation System serial number. 8. Load the “Wash-Concentrate Bead” protocol from the USB stick or from the Rotea protocol download page. 9. ™Now you are ready to load the Gibco™ CTS ... china y rusherkingWebTransport Assays-Just prior to counterflow assay, frozen proteo- liposomes were thawed rapidly in a water bath at room temperature, sonicated for 10 s, diluted with 4 ml of PSDL, and centrifuged at 81,000 X g for 1 h (24). The pellet was suspended in 50 pl of PSDL to obtain a protein and lipid concentration of approximately 1.0 and grand banks performance sweatshirtWebSep 8, 2024 · Application of the competitive counterflow assay to identify novel OATP substrates. To identify OATP1B1 and OATP1B3 substrates, we used the competitive … china youzan investor relationsWebHere’s what researchers using CTS Rotea Counterflow Centrifugation Systems in their labs have to say about the product: “Rotea is exactly what the cell therapies sector has been … grand banks tachometerWebMay 12, 2024 · The activities of GLUT3 variants were examined in the proteoliposome-based counterflow assay. Empty liposome refers to protein-free control. The transport … china youtube channelWebrequirements. The general protocol for these "homogeneous" assays is "add-mix-measure." Some of the assay systems require combining components to create the "Reagent," and some protocols require incubation or agitation steps, but none require removal of buffer or medium from assay wells. The available homogeneous assay … china young mens winter coats factory