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Advanced Western Blotting Solutions for Cancer & Immuno-Oncology

Simple Western Cancer & Immuno-oncology Research applications

What do ProteinSimple’s Advanced Western Blotting Systems Offer Cancer & Immune Therapy Researchers?

ProteinSimple is a pioneer in bringing western blotting into the 21st century with our innovative product lines including our fully automated Simple Western systems and our Single-Cell Western platform.

Simple Western systems provide fully automated, hands-free western blotting analysis on 24 samples per 3 hour run, giving you reproducibility that's spot on, and using less sample in the process (as low as 3 µL sample). With Simple Western assays you can obtain highly quantitative and reproducible results giving you confidence in your data.

Simple Western Systems can help you reveal new insights in cancer & immuno-oncology research and advance the study of proteins, their function and signaling through reliable, quantitative, and automated workflows. Download this publication spotlight for a glimpse of high impact peer-reviewed cancer research with Simple Western tools.

From Your Peers

Krishna C. Yalla, Ph.D., Postdoctoral Research Scientist, Institute of Cancer Sciences, University of Glasgow

“I was able to screen various antibodies in a single run within half a day. This had previously taken weeks and months using traditional Western blotting methods.” - Krishna C. Yalla, Ph.D., Postdoctoral Research Scientist, Institute of Cancer Sciences, University of Glasgow

"Wes is easy to use, convenient, and gives answers quickly. The throughput and system automation is a definite advantage." - Jane Gray, Ph.D., Head of Research Instrumentation, Cancer Research UK Cambridge Institute

Jane Gray, Ph.D., Head of Research Instrumentation, Cancer Research UK Cambridge Institute


Single-Cell Western systems provide the ability to study heterogeneity in your cancer and immuno-oncology samples and to work with low abundance samples (10,000 cells) that cannot be analyzed using flow cytometry. Just probe with your favorite conventional western blot antibodies to measure ~12 proteins per cell using a variety of multiplexing strategies. Our Single-Cell Western technology on Milo unlocks the single-cell proteome to measure more of the proteome than was possible with any other single-cell protein analysis technique.

Single-Cell Western Instrument Applications for Profile Heterogeneity and Cancer Research

Single-Cell Western

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If you’re still doing traditional westerns, our FluorChem Imaging systems also provide robust, reliable and multiplexed images of your gels and blots.

Together, these systems allow for quantitative results that enable identification of specific proteins in complex mixtures and at single cell levels.

FluorChem Imager Instrument Applications for Fluorescence and Chemiluminescence and Cancer Research

FluorChem Imagers

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Study the Tumor Microenvironment

Tumor heterogeneity poses a significant challenge to the development of effective cancer therapies. Further, the tumor microenvironment creates a complex ecosystem of diverse tumor and immune cell types. Methodologies like flow cytometry have limitations such as off-target binding of variants/ isoforms and difficulties in finding validated antibodies. In this application note, Simple Western and Single-Cell Western systems partner to get you critical answers on the immune cell populations found in the tumor microenvironment offering both population level views and single-cell level perspectives on cell subtypes. Request pricing or more information on Simple Westerns and Single-Cell Westerns.

Simple Western Chemiluminescence CD56 and Single-Cell Western CD56 PeakArea Immune Cell Population Test Results

Tumor Landscape Mapping of Chemo-Resistant Metastatic Bladder Cancer with Single-Cell Analysis Tools

Webinar on Tumor Landscape Mapping of Chemo-Resistant Metastatic Bladder Cancer with Single-Cell Analysis Tools

In this webinar, Dr. Kyeung Min Joo, from Sungkyunkwan University School of Medicine, Seoul, South Korea describes their findings from the use of single-cell analysis technologies including Single-Cell Westerns to depict the tumor landscape of a single case of chemo-resistant metastatic, muscle-invasive urothelial bladder cancer (MIUBC).


Drive Your Immuno-Oncology Discovery

The new and rapidly expanding field of immuno-oncology requires high-resolution, quantitative analytical tools at both the population and single-cell level to parse the complex interplay between cancer, immune, and stromal cells. As a result of the increased linear range, small sample size requirements and quantification, Simple Western systems have been involved in breakthrough immunotherapies, including those involving immune checkpoint inhibitors, CAR T-cell therapy, bispecific antibodies, and macrophage switching.

Meanwhile, Single-Cell Western with Milo provides insight into population heterogeneity at the protein level and has been used to study the molecular underpinnings of malignant cell transformation with unprecedented detail. This Scientific Review highlights recent studies that demonstrate the use of Simple Western and Single-Cell Western assays to make breakthroughs in immuno-oncology.


Detect biomarkers in precious, low abundance samples including laser Capture Microdissection (LCM) samples

Laser capture microdissection (LCM) is a powerful technique that has become instrumental to cancer researchers interested in comparing the molecular profiles of tumor tissue to surrounding non-tumor cells. As such, LCM analysis is leading the way for the development of tailored treatment and personalized medicine. Simple Western requires only a few microliters of LCM sample to generate multiple data points. Simple Western also has the sensitivity to analyze low abundance proteins that are difficult to detect by traditional Western blot. And it’s all wrapped up in a simple workflow that minimizes your hands-on time. In this application note, we show two examples of how Simple Western was ideally suited for the analysis of LCM samples. Request pricing or more information on Simple Westerns.

Milo can analyze samples which contain as low as 10,000 single-cells so you no longer have to collect millions of cells to analyze protein expression. The number of cells captured scales with the number of cells loaded so even lower abundance samples are possible. Milo can also characterize highly enriched FACS-sorted cell populations which don't contain enough cells to analyze with other techniques. Request pricing or more information on Single-Cell Westerns.

Characterize Cell Signaling with Quantitative Western Blotting

Quantitative measurements of protein expression are extremely difficult with traditional western blotting due to variability during membrane transfer. Further, simultaneous phospho-protein and total protein measurements on the same sample are challenging on conventional western blots due to loss of protein and non-uniformity during stripping & reprobing. With Simple Westerns, proteins are immobilized after separation, reducing variability and enabling quantitative measurements of protein abundance. Further, multiplexing is easy using multiple detection channels or RePlexing cycles to enable phospho/total measurements in the same capillary. Total protein measurements can also be made on the same sample for more accurate and robust normalization. Request pricing or more information on Simple Westerns.

Quantitative Western Blotting Results for Probe 1 Phospho AKT and Probe 2 Total Protein Detection with Chemiluminescence MCF7 + hlGF1 and MCF7 Control Peak Results


The Milo Single-Cell Western system helps detect discrete cell signaling states by measuring multiple phospho-proteins in individual cells within a population.  Because cells are lysed in Single-Cell Westerns, membrane permeabilization is not required and both intracellular and intranuclear proteins are accessible for antibody probing, making it possible to use phospho- antibodies validated for western blotting and simplifying some challenging intracellular flow assays. This application note demonstrates how Milo adds single cell resolution and heterogeneity information to cell signaling studies traditionally done with conventional western blotting and simplifies challenging phospho-flow assays done with flow cytometry. Request pricing or more information on Single-Cell Westerns.

Detect Epigenetic and Protein Alterations in Single Nuclei

We increasingly understand and appreciate the important role epigenetic alterations play in cancer research including cooperating with genetic alterations to drive the cancer phenotype. In this application note, we discuss method development to use Single-Cell Westerns to investigate histone modifications in single nuclei and understand epigenetic regulation of gene expression at the single-cell level.

H3K27me3 and histone H3 signal distributions (RFU) and co-detection analysis. H3K27me3 was detected in ~68% of cells identified by histone H3 expression.


Oncogenic transformation alters cellular protein expression and metabolism in ways that may be characteristic of the type or stage of cancer. Because of their versatility, FluorChem imagers have been harnessed for applications beyond the Western blot, including DNA gels, colony counting, and imaging Proteome Profile Antibody Arrays from R&D Systems. In this application note, we’ll give you tips on how to image Proteome Profiler Antibody Arrays using chemiluminescent or IR detection on FluorChem M or FluorChem R systems. Together the FluorChem system and Proteome Profile Antibody Arrays provide rapid, sensitive, and economical tool to detect differences in cancer-related proteins between samples.

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