Overview Product Name SDS-PAGE Protein Loading Buffer 5X (Reducing) SKU/Catalog Number AR1112 Form Liquid Size 3mL Contents 10% SDS, 500mM DTT, 50% Glycerol, 250mM Tris-HCL and 0.5% bromophenol blue dye, PH6.8. Description Bosters SDS PAGE Sample Buffer 5X (Reducing) is the most commonly used sample buffer for Sodium Dodecyl Sulfate - Polyacrylamide Gel Electrophoresis (SDS-PAGE) of denatured proteins in the Laemmli SDS-PAGE system. Equivalent GeneCopoeia (Product No. P013) Cite This Product SDS-PAGE Protein Loading Buffer 5X (Reducing) (Boster Biological Technology, Pleasanton CA, USA, Catalog # AR1112) Storage Upon receipt store at -20C. SDS PAGE Sample Buffer 5X (Reducing) is stable for one year. Product is shipped on ice. Assay PrincipleUsing bromophenol blue dye, SDS-PAGE Protein Loading Buffer is a ready-to-use 5X solution. It contains 10% SDS, 500Mm DTT, 50% Glycerol, 500mM Tris-HCL and 0.05% bromophenol blue dye. It can be used for SDS-PAGE protein loading of conventional proteins. It is especially formulated for protein sample preparation to be used in the Laemmli SDS-PAGE system. It is sufficient to prepare 15 mL protein samples. And the whole sample preparation process can be finished within 5 minutes.Assay Protocol1.Equilibrate SDS-PAGE Protein Loading Buffer 5X to room temperature or thaw Loading Buffer in a water bath no higher than 30C.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 Notes1. 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.