The concentrations of purified exosomes were determined using Bradford protein assays (Sangon Biotech, Shanghai, China). Proteins (5 μg) from isolated S. japonicum EVs were separated using precast 4–20% polyacrylamide linear gradient gels (Bio-Rad, Hercules, CA, USA). A pre-stained protein standard (Thermo Scientific, Waltham, MA, USA) was used to track protein migration. After running, gels were stained by silver as previously described46 (link) and scanned using a Bio-Rad Molecular Imager FX system (Bio-Rad).
Polyacrylamide linear gradient gels
Manufactured by Bio-Rad
Sourced in United States
4–20% polyacrylamide linear gradient gels are a type of lab equipment used for protein separation and analysis. They consist of a polyacrylamide gel with a continuous gradient of acrylamide concentration ranging from 4% to 20%. This gradient allows for the separation of a wide range of protein sizes within a single gel.
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2 protocols using polyacrylamide linear gradient gels
1
Quantification and Separation of S. japonicum Extracellular Vesicles
2
Quantifying Schistosoma japonicum EV Proteins
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The protein concentrations of isolated EVs were determined using Bradford protein assays (Sangon Biotech, Shanghai, China). Proteins (1.5 μg) from isolated S. japonicum EVs were separated using precast 4–20% polyacrylamide linear gradient gels (Bio-Rad, Hercules, CA, USA). A pre-stained protein standard (Thermo Scientific, Waltham, MA, USA) was used to track protein migration. The gels were sliver-stained as previously described [8 (link)] and scanned using a Bio-Rad Molecular Imager FX system (Bio-Rad).
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