Wedgewell tris glycine polyacrylamide gels

Manufactured by Thermo Fisher Scientific

12% WedgeWell Tris-Glycine polyacrylamide gels are pre-cast electrophoresis gels used for the separation and analysis of proteins in a sample. These gels have a 12% acrylamide concentration and are based on the Tris-Glycine buffer system.

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Lab products found in correlation

Instantblue coomassie solution, by Abcam (2 mentions) Image studio lite, by LI COR (2 mentions)

Close spelling variants of this product

Tris-glycine polyacrylamide gels Polyacrylamide Tris–Glycine gels Tris-Glycine gels WedgeWell Gel 4–12% polyacrylamide gel Glycine

2 protocols using wedgewell tris glycine polyacrylamide gels

Noc-NBS/parS DNA Crosslinking Kinetics

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A 50 μL mixture of 10 μM Noc (WT/mutants) ± 1 mM NTP ± 1 μM 22-bp NBS/parS dsDNA was assembled in a reaction buffer [10 mM Tris-HCl pH 7.4, 200 mM NaCl, and 1 mM MgCl₂] and was incubated for 10 min at 22°C or for 1, 5, 10, 15, and 30 min at 4°C. Subsequently, BMOE was added to the final concentration of 1 mM, and the reaction was quickly mixed by three pulses of vortexing. SDS-PAGE sample buffer containing 23 mM β-mercaptoethanol was then added immediately to quench the crosslinking reaction. Samples were heated to 50°C for 10 min before being loaded on 12% WedgeWell Tris-Glycine polyacrylamide gels (Thermo Fisher). Each experiment was triplicated. Polyacrylamide gels were stained in an InstantBlue Coomassie solution (Abcam) and band intensity was quantified using Image Studio Lite (LI-COR Biosciences). The crosslinked fractions were averaged, and their standard errors were calculated in Excel.
CTPɣS was custom synthesized either in-house (Rejzek and Le, 2021 (link)) or by Jena Biosciences.

Jalal A.S., Tran N.T., Wu L.J., Ramakrishnan K., Rejzek M., Gobbato G., Stevenson C.E., Lawson D.M., Errington J, & Le T.B. (2021). CTP regulates membrane-binding activity of the nucleoid occlusion protein Noc. Molecular Cell, 81(17), 3623-3636.e6.

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Crosslinking Assay for Noc-NTP-DNA Interactions

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A 50 µL mixture of 10 µM Noc (WT/mutants) ± 1 mM NTP ± 1 µM 22-bp NBS/parS dsDNA was assembled in a reaction buffer [10 mM Tris-HCl pH 7.4, 200 mM NaCl, and 1 mM MgCl2] and was incubated for 10 min at 22 o C or for 1, 5, 10, 15, and 30 min at 4°C. Subsequently, BMOE was added to the final concentration of 1 mM, and the reaction was quickly mixed by three pulses of vortexing. SDS-PAGE sample buffer containing 23 mM β-mercaptoethanol was then added immediately to quench the crosslinking reaction. Samples were heated to 50°C for 10 min before being loaded on 12% WedgeWell Tris-Glycine polyacrylamide gels (Thermo Fisher). Each experiment was triplicated. Polyacrylamide gels were stained in an InstantBlue Coomassie solution (Abcam) and band intensity was quantified using Image Studio Lite (LI-COR Biosciences). The crosslinked fractions were averaged, and their standard errors were calculated in Excel.
CTPɣS was custom synthesized either inhouse 44 (link) or by Jena Biosciences.

Jalal A.S.B., Tran N.T., Wu L.J., Karunakaran R., Rejzek M., Gobbato G., Stevenson C.E.M., Lawson D.M., Errington J., & Le T.B.K. (2021). CTP regulates membrane-binding activity of the nucleoid occlusion protein Noc.

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