The quinolinium salt-based halide-sensitive fluorescence probe N-(ethoxycarbonylmethyl)-6-methoxyquinolium bromide (MQAE; ab145418, Abcam) was used as a measure of chloride ion influx activity13 (link). Following cell culture, the PRNCs were incubated with 5 mM MQAE for 2 h at RT in the dark, and subsequently washed twice with NbActive 4 (BrainBit). Cells were then treated with 50 μM GABA (A2129, Sigma-Aldrich) or 10 nM GABA analog THIP, δ-GABAAR specific agonist8 (link) (T101, Sigma-Aldrich), then fluorescence intensity was consequently measured at 0, 5, 10, and 20 min at RT. For inhibition assay, the cells were pretreated with 1 μM flumazenil (F6300, Sigma-Aldrich) or 1 μM picrotoxin (P1675, Sigma-Aldrich) or PBS (control) for 45 min at RT, and then stimulated with 50 μM GABA for 10 min at RT. Intercellular MQAE is quenched by 10 μM tributyltin chloride (T50202, Sigma-Aldrich) and 10 μM nigericin sodium salt (N7143, Sigma-Aldrich). The fluorescence intensity was measured by the Gemini EX florescence plate reader (Ex/Em = 360/460; Molecular Devices). The kinetic analysis was performed by using GraphPad Prism 6® software.
T50202
Manufactured by Merck Group
T50202 is a laboratory centrifuge designed for general-purpose applications. It has a maximum speed of 4,000 RPM and an adjustable timer. The unit is compact and suitable for benchtop use.
Automatically generated - may contain errors
2 protocols using t50202
1
Chloride Influx Measurement in PRNCs
2
Snail Toxicology: Heavy Metal Impacts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Snails were cultured from hatching to the adulthood in aquaria containing metal-free reconstituted water [20 (link)]. Control and experimental female snails were maintained in 6 L aquaria with reconstituted water [10 (link)] which was changed weekly. Each age group (4, 5, 6 and 7 months old) cultured in reconstituted water was divided before sacrifice in four experimental subgroups that were exposed for 4 weeks to 6 μg/L of TBT [NOECL reported in 26 (link)] as tributyltin chloride (Sigma-Aldrich, T50202) or for six weeks to each of the following compounds, 2 μg/L of Hg as HgCl2 (Sigma-Aldrich, M1136), 10 μg/L of As as Na3AsO4 7H20 (Sigma–Aldrich, S9663), or 30 μg/L of U as UO2CH2COOH (Ted Pella Inc.19481) (Figure 1 , orange lines). These elements were selected because of their potential toxicity to humans and the environment and it were used at the maximum contaminant levels (MCLs) for human consumption allowed by EPA regulations. Exposure conditions were semi static and each compound was added after changing the aquarium water. No snails died during the exposure period. Shell length, PL, PSL, and DI were measured as described above.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!