> Simple Western Assays > Simple Western Size Assays > Simple Western Size-Based Immunoassays
Immunoassays
Approximately 40 nL of sample is loaded into the capillary and the proteins are separated by size as they migrate through a stacking and separation matrix. The separated proteins are then covalently bound to the capillary wall via a proprietary, photoactivated capture chemistry, replacing and eliminating the need to transfer protein to a membrane. Target proteins are identified using a primary antibody and detected using either an HRP-conjugated secondary antibody and chemiluminescent substrate or a fluorescently labeled secondary antibody. The resulting signal is detected and quantitated using Compass for SW software.
The RePlex feature allows you to run two sequential immunoassays within the same capillary to give you rich protein characterization data in a single detection channel. Pair this with size-based or detection channel-based multiplexing strategies and you will be swimming in data!
Available Molecular Weight Ranges & Detection Reagents
The materials needed to run Simple Western immunoassays are sold in modules called Separation Modules and Detection Modules.
Separation Modules
Abby, Wes and Jess Capillaries are run in 13- or 25-capillary cartridges, sold in 2- and 8-cartridge ‘Separation Modules’ that include prefilled assay plates and required reagents.
Cartridges to perform separation by size are available with reagents for 26, 50, 104 or 200 data points
Sally Sue and Peggy Sue Cartridges to perform separation by size are available with reagents for 26, 50, 100, 200 or 480 data points.
Separation Modules
- 2-40 kDa molecular weight range: Small molecular weight proteins are vulnerable to poor resolution and poor retention in traditional western blotting workflows. With Simple Western, you can now detect challengingly small molecular weight proteins like hormones and cytokines with our optimized low molecular weight separation module. Consistently detect and quantify low molecular weight proteins without having to tag them.
- 12-230 kDa molecular weight range: Our most popular range as it covers the the molecular weight range for the majority of proteins.
- 66-440 kDa molecular weight range: Never want to face a ripped gel, overnight transfer, or sky-high CVs for your high molecular weight proteins again? Done! With the 66–440 kDa Master Kit, you can now enjoy all Simple Western has to offer for a greater range of proteins.
Detection Modules
Immunodetection reagents for a variety of species for both chemiluminescence and fluorescence detection are packaged in Detection Modules. Detection modules work with any Simple Western instrument.
Chemiluminescence
Fluorescence
New to Simple Western Assays? Our Simple Western Kit builder will help you design your kit in 5 easy steps! Start now.
