ProteinSimple has launched a total protein assay for its Simple Western product. The Total Protein Master Kit allows users to detect all proteins separated in the assay simultaneously without the need for an antibody. The new kit is good news to researchers who currently go through the whole western blot protocol just to visualize and quantitate an overexpressed protein or evaluate the amount of a protein in a complex cell lysate.
ProteinSimple develops proprietary systems and consumables for a simpler, more quantitative and affordable protein analysis. Their portfolio includes immunoassay-based systems to quantify protein expression and systems that probe the structure and purity of protein-based therapeutics.
Simple Western™ is a protein separation and detection method where the samples are just loaded and no further action is required. Size-based protein separation, immunoprobing, washing, detection and data analysis are automatized. Variability resulting from the manual processes is eliminated. This technology can process 96 samples at once and get quantitative results in 19 hours.
Immunoassay and Total Protein Assay
Simple Western immunoassays take place in a capillary. Samples and reagents are loaded into an assay plate and placed in the device. 40 nL of sample are automatically loaded into the capillary and separated by size as they migrate through a stacking and separation matrix. The separated proteins are then immobilized to the capillary wall via a proprietary, photoactivated capture chemistry. Target proteins are identified using a primary antibody and immunoprobed using an HRP-conjugated secondary antibody and chemiluminescent substrate. The resulting chemiluminescent signal is detected and quantitated.
In the Total Protein Assay, target proteins separated by size are labeled with a biotin reagent and detected by chemiluminescence using Streptavidin-HRP. At the end of the run, you’ve got relative quantitation for your protein of interest in total protein in the sample.