SDS-PAGE of each TMP sample was performed by using a 5.0% stacking gel and a 10.0% running gel using a Mini-Protean II cell electrophoresis system (Bio-Rad Laboratories, Richmond, CA, USA) according to the method of Liu et al. (2014) (link) under reducing (electrophoresis samples prepared with β-mercaptoethanol) or non-reducing (electrophoresis samples prepared without β-mercaptoethanol) conditions with some modifications. Briefly, 1.0 mL of each TMP solution (2.0 mg/mL) was mixed with 1.0 mL of the sample buffer (containing 4% SDS, 20% glycerol, with or without 10% β-mercaptoethanol, and 0.125 M Tris-HCl, pH 6.8), and heated to 100 °C for 5 min, and then centrifuged at 1800 g to remove any particulates. Aliquots of 20 μL of the supernatants were loaded into each well on the gel. A molecular weight (MW) standard, composed of a cocktail of proteins (from 11 kDa to 245 kDa) (TaKaRa Biotechnology Co., Ltd. Dalian, China) was also run.
Mini protean 2 cell electrophoresis system
Manufactured by Bio-Rad
Sourced in United States
The Mini-Protean II cell electrophoresis system is a laboratory equipment designed for performing gel electrophoresis. It provides a controlled environment for the separation and analysis of macromolecules such as proteins or nucleic acids.
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Lab products found in correlation
2 protocols using mini protean 2 cell electrophoresis system
1
SDS-PAGE Analysis of Protein Samples
2
Protein Quantification and SDS-PAGE Analysis
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Protein concentrations in the EVs and WCP samples were determined by Quick StartTM Bradford protein microplate standard assay (Bio-Rad, USA). For protein separation, the samples were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using the mini-PROTEAN II cell electrophoresis system (Bio-Rad, USA). The proteins were denatured in 2× loading buffer at 100°C for 5 minutes, followed by centrifugation at 5000 ×g for 5 minutes. 20 μl of proteins loaded in each well of the gel were separated on 12% SDS-PAGE at a constant 120 V. After the run was completed, protein bands were detected using silver stain. Gel images were visualized in G: Box Imaging System (Syngene, India). Protein banding patterns and molecular weights of the bands were determined using GeneSys tools software.
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