Protein samples were prepared for Western blot analysis as described previously [14 (link)]. Protein concentrations were quantified using the Bio-Rad protein assay (Bio-Rad, Hercules, CA, USA) with bovine serum albumin (BSA) as a standard. Thirty micrograms of protein were separated on Mini-PROTEAN® TGX™ precast 4–15% gradient gels (Bio-Rad) and transferred to nitrocellulose membranes using the Trans-Blot Turbo transfer system, (Bio-Rad). Primary antibody details and dilutions are listed in Supplementary Table S2 and appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies (1:2000; P0448, DAKO) were used for detection. Proteins were detected using the Pierce ECL Western blotting kit (Thermo Fisher Scientific). Images were captured using the ChemiDoc XRS system and Image Lab version 3.0 (Bio-Rad).
Mini protean tgx precast 4 15 gradient gels
Manufactured by Bio-Rad
Sourced in United States
The Mini-PROTEAN® TGX™ precast 4–15% gradient gels are a type of electrophoresis gel used for the separation and analysis of proteins. These gels feature a 4–15% gradient of polyacrylamide, which allows for the separation of a wide range of protein molecular weights in a single run.
Automatically generated - may contain errors
Lab products found in correlation
Image lab version 3, by Bio-Rad (3 mentions)
Pierce ecl western blotting kit, by Thermo Fisher Scientific (3 mentions)
Chemidoc xrs system, by Bio-Rad (3 mentions)
Trans blot turbo transfer system, by Bio-Rad (3 mentions)
Protein assay, by Bio-Rad (3 mentions)
P0448, by Agilent Technologies (2 mentions)
α sma, by Merck Group (1 mentions)
Collagen 1, by Southern Biotech (1 mentions)
3 protocols using mini protean tgx precast 4 15 gradient gels
1
Western Blot Analysis of Protein Samples
2
Western Blot Analysis of Protein Samples
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Protein samples were prepared for Western blot analysis as described previously [10 (link)]. Protein concentrations were quantified using the Bio-Rad protein assay (Bio-Rad, Hercules, CA, USA) with bovine serum albumin (BSA) as a standard. Samples totaling 30 µg protein were separated on Mini-PROTEAN® TGX™ precast 4–15% gradient gels (Bio-Rad) and transferred to nitrocellulose membranes using the Trans-Blot Turbo transfer system (Bio-Rad, Hercules, CA, USA). Primary antibodies used were α-SMA (mouse monoclonal, Sigma Aldrich, A5228, 1/1000), Collagen-I (goat polyclonal, Southern Biotech, Homewood, AL, USA, 1310-01, 1/1000) and GAPDH (rabbit polyclonal, Calbiochem, CB1001, 1/10,000), and appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies (1:2000; P0448, DAKO, Glostrup, Denmark) were used for detection. Proteins were detected using the Pierce ECL Western blotting kit (Thermo Fisher). Images were captured using the chemidoc XRS system and Image Lab version 3.0 (Bio-Rad).
3
Western Blot Analysis of Hepatic Proteins
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Protein samples were prepared for Western blot analysis as described previously [26] (link). Protein concentrations were quantified using the Bio-Rad protein assay (Bio-Rad, Hercules, CA, USA) with bovine serum albumin (BSA) as a standard. Equal amounts of protein (5-20 μg) were separated on Mini-PROTEAN® TGX™ precast 4-15% gradient gels (Bio-Rad, Hercules, CA, USA) and transferred to nitrocellulose membranes using the Trans-Blot Turbo transfer system, (Bio-Rad). Primary antibodies (against HSL, pHSL, αSMA, Collagen1A1, LRAT and GAPDH; details and dilutions are listed in Supplementary Table S2) and appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies (1:2000; P0448, DAKO) were used for detection. Proteins were detected using the Pierce ECL Western blotting kit (ThermoFisher Scientific). Images were captured using the chemidoc XRS system and Image Lab version 3.0 (Bio-Rad). The intensity of bands was quantified using ImageJ version 1.51 (NIH, USA). The Ponceau-S staining of the membrane is included as loading control.
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