SDS-PAGE Protein Loading Buffer 5X (Reducing)

Li-Juan Nie 1 2, Zhe Cheng 1, Yi-Xian He 1, Qian-Hua Yan 1, Yao-Huan Sun 1, Xin-Yi Yang 1, Jie Tian 1, Peng-Fei Zhu 1, Jiang-Yi Yu 1, Hui-Ping Zhou 3, Xi-Qiao Zhou Role of duodenal mucosal resurfacing in controlling diabetes in rats World J Diabetes, (2025)

Assay Protocol 1. Equilibrate SDS-PAGE Protein Loading Buffer 5X to room temperature or thaw Loading Buffer in a water bath no higher than 30°C. 2. Mix one volume of SDS-PAGE Protein Loading Buffer 5X with four volume of protein sample (i.e. add 4mL protein sample into 1 mL Loading Buffer) . 3. Boil sample for 3-5 min. Note: If semitransparent viscous substance remains after boiling, boil sample for another 5-10 min or add 1X SDS-PAGE Protein Loading Buffer which is diluted from 5X SDS-PAGE Protein Loading Buffer to the sample and then boil for another 3-5 min. 4. Allow sample to cool to room temperature. 5. Load sample into the wells of the SDS-PAGE gel and begin electrophoresis. 6. Stop electrophoresis when bromophenol blue dye front reaches to the bottom of the gel. General Notes 1. SDS-PAGE Protein Loading Buffer 5X contains DTT which has a slightly irritating odor. 2. SDS-PAGE Protein Loading Buffer 5X contains DTT which has a slightly irritating odor. 3. Please wear the lab coat and disposable gloves and operate in a laboratory hood.