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High Sensitivity Multiplexing and Total Protein Normalization

The understanding of cell signaling pathways has significant potential to advance therapies in cancer and regenerative medicine. Simple Western™ from ProteinSimple, a Bio-Techne brand, is a capillary-based automated immunoassay that separates proteins by Charge or Size for specific and sensitive protein characterization using conventional Western blot antibodies.

  • Unlike the traditional Western blot, Simple Western assays generate reproducible and fully quantifiable data in as little as 3 hours. And it's all automated.
  • RePlex™ on Simple Western performs two sequential immunoassays in the same capillary, allowing users to quantify the relative phospho-isoforms to total protein isoforms of any target without the need to strip & reprobe.
  • Take multiplexing even further with chemiluminescence, infrared (IR), and near-infrared (NIR) detection channels.
  • Stop the use of housekeeping controls, which can fluctuate in response to cell stimuli with misleading results. Instead, use built-in Total Protein Detection for more accurate quantification of your phospho/pan protein pair in response to cell stimuli.
RePlex on Jess quantifies phospho and pan AKT expression with total protein normalization
Automated multiplex detection of phospho/pan AKT and total protein normalization with RePlex.

Analyze an Entire Cell Signaling Pathway Overnight

Cell signaling analysis can be performed in each of your samples in a single experiment. Simple Western can analyze up to 96 samples in a single overnight run using only 5 μL of sample, which is ideal for gaining a comprehensive overview of many targets together with their phosphorylated isoforms in a complex cell signaling pathway in a single experiment. Follow our sample prep workflow for large sample numbers, then load your samples in Peggy Sue, Sally Sue, or NanoPro 1000 and let Simple Western take care of the rest!

The automated high-throughput Simple Western assay can analyze up to 96 samples overnight, enabling the analysis of full signaling pathways in one run.

Unraveling the Cell Signaling Pathway in Stem Cells

Learn how high sensitivity multiplex assays on Simple Western monitor iPSC differentiation into three primary germ layers of human embryos using GMP proteins and GMP small molecules. Reveal and quantify changes in the expression of pluripotent factors and cell signaling proteins that regulate stem cell differentiation with only 3 μL of starting material. Get unrivaled multiplex flexibility, sensitivity, and reproducibility for accurate protein expression measurements that scale with manufacturing workflows for stem cell therapy and regenerative medicine.

stem cells in culture at Bio-Techne

High-Resolution Protein Isoform Analysis with Simple Western Charge

Simple Western charge separation diagram

What is Simple Western Charge?

This capillary isoelectric focusing (cIEF) immunoassay has many advantages over slab gel IEF and Western blot.

eBook on Simple Western Charge

Simple Western Charge Protocols

Detect and quantify nearly all protein isoforms with high sensitivity and resolution.

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Detailed ERK1/2 Characterization

Resolve and quantify unphosphorylated, mono, and dual-phosphorylated isoforms of ERK1/2.

Single-Cell Western Blot Analysis

Single-Cell Westerns on Milo™ perform Western analysis on approximately 1000 individual cells to provide insight into population heterogeneity. Like Simple Western, Single-Cell Western uses conventional Western blot antibodies and is capable of resolving phosphorylated isoforms that are critical for understanding the activity of cell signaling networks. Taken together with Simple Western, these advanced Western blotting solutions offer protein characterization at both a population-level and single-cell resolution.

Milo Single Cell Western Blot Platform Hero Image

Uncover Phospho-Protein Heterogeneity with Milo

Single-Cell Westerns on Milo perform measurements that are difficult or impossible to make with conventional westerns or flow cytometry. Learn how Milo can simultaneously quantify phospho- and total protein expression in each individual cell and quantify how the extent of phosphorylation varies across stimulated and unstimulated cell populations.

Customer Success Stories

Simple Western Applications

immune cells attack cancer cell



Transform your cancer & immuno-oncology research by utilizing next-generation Western blotting tools to give you richer views into your samples.

Solutions for Cell Signaling and Signal Pathways Analysis

Cell Signaling


Analyze entire signaling pathways with high sensitivity multiplexing and total protein normalization.

Testing for Lentivirus and AAVs

Cell and Gene Therapy


Next-generation analytical instrumentation to help improve and accelerate your cell and gene therapy workflows.

Advances in Targeted Protein Degradation

Targeted Degradation


Boost your screening studies using high throughput & hands-free Simple Western systems for quantitative analysis.

Biosimilar Antibodies Teaser Image

Pharmacokinetic Analysis


High-throughput pharmacokinetic analysis with quantitative, high-sensitivity assays directly in complex lysates and tissue homogenates. 




A validated workflow for protein analysis in extracellular vesicle samples from the leaders in exosome isolation and high sensitivity Western blotting solutions.

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Vaccine Development


Improve reproducibility, quantitation, and time to market for your vaccine candidates.

Neuroscience App image for Ella



Increase your sensitivity and multiplexing capabilities with neural samples using automated Western blotting systems.

Simple Plex HEK-HCP Assay

Bioprocess Impurities


Screen for processing-related impurities and host-cell related contaminants. Quantify empty/full capsid content of viral vectors like AAVs.


SARS-CoV-2 Research


Reduce your lab time while accelerating your COVID‑19 research. Set up, leave and remotely review your data in just 3 hours!

SARS-CoV-2 Serology

SARS-CoV-2 Serology Assay


Rapid quantitative characterization of human IgG antibodies in serum or plasma reactive against multiple SARS-CoV-2 antigens.