ProteinSimple Instruments Ella, Maurice, and Jess
Simple Western Assays Utilizing Size-Based Separation

> Simple Western Assays > Simple Western Size Assays

A Simple Western size assay is an automated Western blot — no gels, no transfer devices, no blots, no film and no manual analysis! Just load your samples in a Simple Western system and press start—it's a complete, walk-away solution for molecular weight-based protein separation.

Simple Western size-based immunoassays and total protein assays are super straightforward. The initial sample preparation for all size-based assays is very similar to a traditional Western Blot where the sample is prepared in an SDS sample buffer that contains a reducing agent. Simple Western just requires additional fluorescent standard mix used to determine the molecular weight of your protein of interest. Samples are then heat-denatured and pipetted onto a Simple Western plate along with required detection reagents. From there, just load your capillary cartridge and sample plate and push run. That’s it! You can design your experiment to probe for your protein target(s) of interest in an immunoassay and/or use one of several total protein assay chemistries to visualize all the protein bands in your sample.

Multiplexed Western Blotting Assays

All size-based Simple Western immunoassays offer the flexibility to multiplex and detect multiple proteins per sample, giving you more data from the same sample. Plus, you can combine single or multiplexed immunoassays with total protein assays to normalize your immunoassay protein expression data with total protein expression on each sample. Just load your samples and off to the races you go!

Singleplex Assays Multiplex Assays
or Fluorescence
or Total Protein
Size based multiplexing
2+ immunoassays
in 1 detection channel
Detection channel
based multiplexing


immunoassay 1
+ Chemiluminescence
immunoassay 2
immunoassay 1
+ Fluorescence
immunoassay 2
+ Fluorescence
+ Total protein assay using the Protein Normalization module
Probe 1: Fluorescence/
+ Probe 2: Fluorescence/
Total protein assay

Advantages of Increasing Assay Plex

  • More data points per sample
  • Multiple protein detection
  • Faster time to data
  • Lower cost / sample
    Superplexing All the advantages of multiplexing, plus:
  • Chemi, fluorescence, and total protein normalization (PN) in the same capillary
    RePlexing All the advantages of Superplexing, plus:
  • Two independent immunoprobes in the same capillary
  • Greater flexibility in antibody selection
  • Higher sensitivity protein normalization
  • Quantify phosphorylated target and total target

Learn more about the various multiplexing strategies available including Superplexing and RePlexing.

Learn More about Multiplexing

Applications Specific Kit: SARS-CoV-2 Multi-Antigen Serology Module

The SARS-CoV-2 Multi-Antigen Serology Module is designed for simultaneous detection of human serum IgG reactivity with 5 viral antigens of interest for characterization of the COVID-19 immune response, including the nucleocapsid, the full-length spike protein, as well the spike protein S2, S1, and S1 RBD subunits. Analyze up to 24 samples within one 3-hour run to advance understanding of the human immune response to SARS-CoV-2! Learn more about the benefits of the SARS-CoV-2 serology assay.

Proof of concept studies have also shown that SARS-CoV-2 Multi-Antigen Serology Module for Jess/Abby/Wes can be used to monitor the COVID-19 immune response in animal systems such as mice when the appropriate secondary antibody is chosen. Learn more about the benefits of the SARS-CoV-2 serology assay.

Abby Instrument Used for Monitoring COVID-19 Immune Response with the SARS-CoV-2 Multi-Antigen Serology Module

SARS-CoV-2 Multi-Antigen Serology Module for Jess and Wes Simple Western Instruments

Jess Instrument Used for Monitoring COVID-19 Immune Response with the SARS-CoV-2 Multi-Antigen Serology Module