Sds polyacrylamide bis tris gels

Manufactured by Thermo Fisher Scientific

SDS–polyacrylamide Bis-Tris gels are electrophoresis gels used to separate proteins based on their molecular weight. They utilize a Bis-Tris buffer system and sodium dodecyl sulfate (SDS) to denature proteins, allowing for their separation by size.

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Bis-Tris SDS-polyacrylamide gel 4–12% Bis-Tris polyacrylamide gel SDS Bis-Tris gel SDS-polyacrylamide gels Bis-Tris gels 4–12% polyacrylamide gel

2 protocols using sds polyacrylamide bis tris gels

Ubiquitin Topoisomers Biochemical Characterization

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Ubiquitin monomer, BSA, Tris, Triton X-100 and DTT, trimethyl-psoralen (TMP), hydroxyurea, angelicin and BrdU were purchased from Sigma-Aldrich. Di-ubiquitin topoisomers (linear, K6-, K11-, K27-, K29-, K33-, K48- and K63-linked ubiquitin–ubiquitin) were purchased from Boston Biochem. Complete protease inhibitors cocktail tablets were obtained from Roche. Tween-20, Colloidal blue staining kit and precast SDS–polyacrylamide Bis-Tris gels (Invitrogen). MMC was from Duchefa.
All recombinant proteins, plasmids and antibodies generated for the present study are available upon request and described in further detail on our reagents website (https://mrcppureagents.dundee.ac.uk/).

Perez-Oliva A.B., Lachaud C., Szyniarowski P., Muñoz I., Macartney T., Hickson I., Rouse J, & Alessi D.R. (2014). USP45 deubiquitylase controls ERCC1–XPF endonuclease-mediated DNA damage responses. The EMBO Journal, 34(3), 326-343.

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Protein Quantification and Silencing

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Antibodies for β-actin were obtained from Abcam, and anti-phosphorylated (p)NKCC2 was a gift from Dr. Kerim Mutig. Lipofectamine reagents, TRIzol, and SDS polyacrylamide bis-Tris gels were from Invitrogen. Silencing of TNF mRNA was accomplished using the lentiviral vector psiLv-U6 (GeneCopoeia) and the lentivirus purification kit from Takara Bio USA. Recombinant mouse TNF was obtained from PharMingen. Polyvinylidene difluoride membranes were obtained from Amersham. The Nonidet P-40 lysis buffer contained protease inhibitors (Roche Diagnostics) for anti-pNKCC2 analysis. Tissue culture media were obtained from Life Technologies (Grand Island, NY). The luciferase assay kit was from Promega (Madison, WI). All other chemical reagents were purchased from Sigma.

Hao S., Zhao H., Hao D.H, & Ferreri N.R. (2022). MicroRNA-195a-5p regulates blood pressure by inhibiting NKCC2A. Hypertension (Dallas, Tex. : 1979), 80(2), 426-439.

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