Western blot was adopted from studies performed by Odegaard and colleagues (Odegaard et al., 2020a (link)). Brain and liver lysates, and synaptosomes from each animal were loaded into SDS PAGE 10 well gels (Invitrogen, Waltham, MA, USA) under reducing conditions, followed by transfer to a nitrocellulose membrane using iBlot2 (Invitrogen) and immunodetection. Concentrated Superblock was used to block nonspecific binding (Thermo Fisher Scientific, Waltham, MA, USA). After blocking, membranes were incubated overnight at 4°C with a primary antibody. Primary and secondary antibody dilutions were done according to the manufacturer’s suggestion and are shown inTable 1 .. Blots were developed using Azure cSeries Imager (Azure Biosystems, Dublin, CA, USA) with Super-Signal West Pico Chemiluminescent Substrate (Thermo Fischer Scientific).
Sds page 10 well gels
Manufactured by Thermo Fisher Scientific
Sourced in United States
SDS PAGE 10 well gels are electrophoresis gels used for the separation and analysis of proteins. They provide 10 sample wells for loading protein samples. The gels are designed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) technique, which is a standard method for protein separation based on molecular weight.
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2 protocols using sds page 10 well gels
1
Western Blot for Brain, Liver, and Synaptosome Protein Analysis
2
Western Blot Analysis of Brain, Liver, and Synaptosomes
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Western blot was adopted from studies performed by Odegaard and colleagues (Odegaard et al., 2020a) . Brain and liver lysates, and synaptosomes from each animal were loaded into SDS PAGE 10 well gels (Invitrogen, Waltham, MA, USA) under reducing conditions, followed by transfer to a nitrocellulose membrane using iBlot2 (Invitrogen) and immunodetection. Concentrated Superblock was used to block nonspecific binding (Thermo Fisher Scientific, Waltham, MA, USA). After blocking, membranes were incubated overnight at 4°C with a primary antibody. Primary and secondary antibody dilutions were done according to the manufacturer's suggestion and are shown inTable 1.. Blots were developed using Azure cSeries Imager (Azure Biosystems, Dublin, CA, USA) with Super-Signal West Pico Chemiluminescent Substrate (Thermo Fischer Scientific).
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