2024 Scrape loading assay

Scrape loading assay

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August undefined, 2024

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Gap junction mediated intercellular transfer of miRNAs assessed …

WebFeb 1, 1987 · Scrape-loading has been effectively utilized to introduce macromolecules into cells in culture by inducing a transient tear in the plasma membrane without affecting cell viability or colony-forming ability [27]. The tracer dye Lucifer yellow (MW 457.2) is an intensely fluorescent 4-ammophthali- mide with a high quantum yield of about 0.25 [28, 29]. 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% Lucifer Yellow (10 ml for 6 well plates, 0.1 g LY/10 ml PBS) PBS; 2-4% paraformaldehyde For small cover slips: Move the cover slip from the 24 well to a 3 cm plate with media. fizik off road shoes

Identification of Cx45 as a Major Component of GJs in HeLa Cells

WebDownload scientific diagram Gap junction mediated intercellular transfer of miRNAs assessed by scrape-loading assay. ( a–c ) Intercellular diffusion of fluorescence-tagged miRNA (miR-F) and ... WebJul 4, 2024 · Scrape loading and dye-uptake assays. For scrape loading assay to determine gap junctional intercellular communication, after scrapes were made with a blade, cells were incubated with 0.5 mg/ml Neurobiotin (Vector Laboratories, 1120) for 10 min followed by a normal media incubation of 20 min. For dye uptake assay to investigate the hemichannel ... 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. can non military get usaa renters insurance

Advantages and limitations of commonly used methods …

WebSep 1, 2003 · For LY scrape-load assays,quantification was based on measuring the integrated fluorescence intensity,at the scrape site, over an equivalent area in both control and experimental conditions. The degree of significance of variations between control and experimental values was assessed by ANOVA uni-variant test using the Graph Pad Prism … 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% … can non military use navy federalNational Center for Biotechnology Information cannon mini mag electric downrigger manuals

"WebJan 9, 2024 · Following the scrape-loading of a low MW ... The LY dye uptake assay relied on the ability of this dye present in the extracellular milieu to cross the plasma membrane and became concentrated ... " - Scrape loading assay

Scrape loading assay

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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 … WebMay 16, 2024 · The scrape-loading technique can be effectively used as a tool to determine the qualitative presence or absence of GJIC. Limitations The approach is inappropriate to …

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WebWe conducted the I-YFP GJIC assay in HeLa cells, which revealed a weak level of GJIC that could not be detected by the Lucifer yellow scrape-loading assay. The mRNA expression of GJB5 (Cx31.1), GJA1 (Cx43), and GJC1 (Cx45) was detected in … 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 monitoring of its transfer into contiguous cells. ... Glial cell viability was measured by MTT assay. Immunocytochemistry was performed to analyze the microglial activation state. Western …

WebDec 25, 2016 · The assay involves pre-loading cells with a fluorescent dye by briefly perturbing the cell membrane with a scalpel blade through a population of cells. The … WebMay 1, 2000 · Several in vitro assays have been developed to determine the effect of chemicals on gap junction communication in cultured cells. The scrape loading dye transfer technique is based on studying the transfer of the fluorescent dye Lucifer Yellow in cells where the dye is loaded through a cut in the cell monolayer.

WebJun 1, 2000 · The scrape-loading dye transfer assay allows simultaneous monitoring of dye coupling in a large population of cells; its suitability for the assessment of gap junction-mediated intercellular...

WebMar 1, 1987 · The scrape loading-dye transfer (SL-DT) technique is a simple assay for the functional evaluation of GJIC in various in vitro cultured mammalian cells and …

WebScratch loading is a variation of the scrape loading technique (Swanson and McNeil, 1987). An artificial wound is made on a cell monolayer with a sharp instrument in the presence of … cannon motor companyWebJan 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 junction-permeable fluorescent dye, typically Lucifer Yellow (457 Da, negatively charged), which is introduced into adherent cells typically grown in a Petri dish. cannon mmc plus 32 driver window 10WebMay 27, 2024 · The scrape loading/dye transfer assay is a commonly used approach to study intercellular coupling via functional gap junction channels (el-Fouly et al., 1987). Gap junction intercellular communication (GJIC) was evaluated in undifferentiated ROSA26 and Cx43 KD IDG-SW3 cells. Cells were grown to reach confluence before performing the … fiziko fitness sector 23WebAssessment of GJ Communication in hESC Using a Carboxyfluorescein Scrape-Loading Assay (A-F) H1 DE monolayers (day 3) were incubated with rhodamine dextran (Rhod DX) and carboxyfluorescein (CF)... fizik of 4am ane seamWebProtein Extraction Protocol Steps. Discard the medium in culture dishes with cells and wash the cells using ice-cold PBS. Discard the PBS, add ice-cold lysis buffer. Scrape the cells using cold plastic cell scraper. Collect the cells in microcentrifuge tubes. Agitate the contents in microcentrifuge tubes for 30 min at 4 °C. fizik leather bar tapeWebJan 27, 2016 · The scrape-loading assay also showed that there was no intercellular diffusion of miR-F in the Cx26 R75W cell line ; the loaded miR-F was restricted at the scrape edge as shown in the Cx-null cell ... cannon mine coffeeWebJan 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 … cannon mortuary cottonwood heights