Scrape-loading dye transfer assay
WebSep 18, 2014 · Scrape-loading/dye transfer assay for gap junctional intercellular communication Solutions needed: HBSS + 1% BSA (50 ml; 0.5 g BSA/HBSS) PBS + 1% … WebOne of these assays, the scrape-loading dye transfer (SLIDT) technique, is based on monitoring the transfer of the fluorescent dye Lucifer yellow from one cell into adjacent cells via functional gap junctions. The objective of our study was to evaluate and compare various approaches for quantifying results obtained with the SL/DT technique.
Scrape-loading dye transfer assay
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WebSep 3, 2024 · MC3T3-E1 cell line were cultured in a 25 cm2flask containing 3-4 ml α-MEM with 10% fetal bovine serum at 37 °C in a humidified atmosphere of 5% CO2in the air. FGF7 treatment Both primary osteoblasts and MC3T3-E1 cell line were seeded onto six-well plates at 5 × 105cells per well (85%~95% confluence). WebJul 1, 2008 · The scrape loading and dye transfer (SL/DT) method is a functional assay widely used to evaluate the level of the intercellular communication via gap junction. Initially, the evaluation of the degree of communication was done by counting the rows of fluorescent cells from the scrape.
WebFigure 7 Utility of the scrape loading/dye transfer assay to detect a toxin/toxicant s ability to modulate gap junctional intercellular communication. The last two images (6ngmr TPA) illustrate both the epifluorescent and phase images, respectively, of the same cells to show that the complete inhibition of GJIC was not associated with the death ... WebJan 20, 2024 · The commonly used technique for in vitro assessment of GJIC is so-called scrape loading-dye transfer assay (SL-DT) using a membrane-impermeable and gap …
WebJan 24, 2013 · The function of gap junctional intercellular communication (GJIC) was assessed by scrape loading dye transfer assay as previously described. Cells cultured in 35 mm dishes were incubated in preheated (37 °C ) 1% Lucifer Yellow dye solution (dissolved in Ca 2+ and Mg 2+ free PBS) after they were rinsed with warm PBS (37 °C ). WebThe GJIC was investigated by a scrape-loading dye transfer assay. Expression of Cx43 and K3 keratin as well as BrdU-retaining nuclei were also ana-lyzed after xenotransplantation in nude mice. Results: Limbal epithelium showed mean±SD 12.4% ±14.5%positiveunitsof Cx43expressionandalowBrdU
WebDec 22, 2024 · The scrape-loading dye transfer assay adapted from (Babica et al. 2016) was used to assess gap-junction intercellular communication within an EB. A clean linear cut was made across an individual EB plated on a glass coverslip using a curved surgical scalpel blade. The EB was then loaded with lucifer yellow fluorescent dye added into the culture ...
WebFeb 1, 1987 · It employs scrape-loading to introduce a low molecular weight (MW) fluorescent dye, Lucifer yellow CH (MW 457.2) into cells in culture and allows the … how to overhaul your lifeWebMar 8, 2024 · In scrape-loaded astrocytes, the dye traveled through functional gap junctions into adjacent gap junction–coupled astrocytes that avoided the scrape-loading. Astrocytes were pretreated with MHC1 (66.7 nM) or a known gap junction blocker, carbenoxolone (CBX) (100 μM), for either 3 or 24 hours at 37°C prior to the gap junction coupling assay. how to overhaul lawn mower engineWebAug 1, 2024 · The SL-DT assay can serve a broad range of in vitro pharmacological and toxicological needs, and can be potentially adapted for high throughput set-up systems … mx 9900 chipperWebA scrape loading/dye transfer assay was used to assess the effects of the chemicals on GJIC. In this assay, a scratch is made in a confluent cell layer in the presence of a Lucifer … how to overhaul your bodyWebMar 7, 2024 · Scrape-loading dye transfer in vitro. Gap junction-mediated intercellular coupling was evaluated by transfer of lucifer yellow dye in a scrape-loading assay as described previously with minor modifications . Briefly, astrocytes were seeded on 12-well culture plates and incubated at 37 °C and 5% humidified atmosphere. mx 950 editor softwareWebFeb 28, 2011 · An innovative adaptation of traditional, fluorescence microscopy-based scrape loading-dye transfer (SL-DT) assay is presented, which has been optimized to simultaneously evaluate GJIC, cell density and viability, which could be included into in vitro screening and assessment of pharmacologically and toxicologically relevant compounds. … mx 915 pin pad troubleshootingWebMar 1, 1987 · Scrape loading/dye transfer method (SL/DT) was used to analyse gap junction activity of cells, as previously reported [29]. Myocytes and macrophages were seeded in … how to overhaul your diet