Cell culture reagents (DMEM, FBS, PenStrep, and trypsin-EDTA), as well as western blot reagents include NuPAGE 4–12% Bis-Tris Gel and MOPS SDS Running Buffer (20X) were from Life-Technologies (Grand Island, NY). NAO and DAPI were from Molecular Probes (Eugene, Oregon). DCFH, cytochrome c, n-Dodecyl-beta-maltoside, and DTT were from Sigma (St. Louis, MO). 5-Aminoimidazole-4-carboxamide riboside (AICAR) was from Cayman chemical company (Ann Arbor, MI). The antibodies for AMPK (AMP-activated protein kinase) and pAMPK (rabbit monocloncal anti-Phospho-Thr172-AMPKα), PGC1α, SIRT1, phospho-Ser47-SIRT1 (p-SIRT1) were from Cell Signaling Technology (Beverly, MA). Antibodies for SURF1 and Actin were from Santa Cruz Biotechnology (Dallas, TX). Protease inhibitors, phosphatase inhibitors, and RIPA buffer with EDTA were from Boston BioProducts (Ashland, MA). Pierce ECL Western blotting Substrate and Restore Western blot Stripping Buffer were from Thermo Scientific (Rockford, IL). Quick Start Bradford Dye Reagent for protein quantification was from Bio-Rad (Hercules, CA).
Mops sds running buffer 20x
Manufactured by Thermo Fisher Scientific
MOPS SDS Running Buffer (20X) is a concentrated electrophoresis buffer solution used for separating proteins in polyacrylamide gel electrophoresis (PAGE) experiments. It is a 20X concentrated formulation that requires dilution prior to use.
Automatically generated - may contain errors
Lab products found in correlation
Nupage 4 12 bis tris gel, by Thermo Fisher Scientific (2 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Phosphatase inhibitor, by Boston BioProducts (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Dithiothreitol (dtt), by Merck Group (1 mentions)
Restore western blot stripping buffer, by Thermo Fisher Scientific (1 mentions)
Cytochrome c, by Merck Group (1 mentions)
Sirt1, by Cell Signaling Technology (1 mentions)
Protease inhibitors, by Boston BioProducts (1 mentions)
Qubit protein assay kit, by Thermo Fisher Scientific (1 mentions)
Qubit assay tube, by Thermo Fisher Scientific (1 mentions)
Chemidoc mp imaging system, by Bio-Rad (1 mentions)
Qubit fluorometer, by Thermo Fisher Scientific (1 mentions)
2 protocols using mops sds running buffer 20x
1
Western Blot Analysis of Metabolic Regulators
2
Protein Quantification and Separation in EVs
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Protein concentrations in EVs samples were determined using the Qubit Protein Assay Kit (Thermo Fisher Scientific). Measuring was done using the Qubit Fluorometer (Thermo Fisher Scientific). This assay was performed at room temperature using specific Qubit assay tubes (Cat. no. Q32856).
For protein separation, the samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using the Mini Gel Tank electrophoresis system (Invitrogen). Proteins were denatured in a loading buffer (1X LDS sample buffer + β -mercaptoethanol) at 98 °C for 5 min. Samples were loaded and separated on a NuPAGE 4–12% Bis–Tris Gel (Invitrogen) using 1X MOPS SDS Running Buffer 20X (Life technologies) at a constant voltage of 110 V and for 1:30 min. After the run was completed, several bands were identified. Gel images were visualized in the ChemiDoc MP Imaging System (Bio-Rad).
For protein separation, the samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using the Mini Gel Tank electrophoresis system (Invitrogen). Proteins were denatured in a loading buffer (1X LDS sample buffer + β -mercaptoethanol) at 98 °C for 5 min. Samples were loaded and separated on a NuPAGE 4–12% Bis–Tris Gel (Invitrogen) using 1X MOPS SDS Running Buffer 20X (Life technologies) at a constant voltage of 110 V and for 1:30 min. After the run was completed, several bands were identified. Gel images were visualized in the ChemiDoc MP Imaging System (Bio-Rad).
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