On This Page: Customer Success Stories | Protein Size Matters | Types of Protein Size Assays | References
Protein Size Assays on Simple Western
Add a 3rd Dimension to Your Immunoassay
As a capillary electrophoresis immunoassay platform, Simple Western™ provides sensitive, accurate, and reproducible quantitative immunoassay read-outs while also providing molecular weight (MW) information enabled by electrophoretic protein separation. While ELISA can only measure antibody binding as you vary experimental conditions (i.e., 2-dimensional data), Simple Western measures antibody binding activity and provides MW size information as you vary experimental conditions, adding a 3rd dimension to your immunoassay.
Download our Scientific Reviews Summary to learn why adding MW protein size separation to your immunoassay data matters.
Customer Success Stories
Director of Neuroscience Center of Excellence, LSUHSC
Senior Product Scientist, Cell Signaling Technology (CST)
Postdoctoral Fellow, University of Texas Health Science Cent
Postdoctoral Research Scientist, ICS, University of Glasgow
Protein Size Matters
Specificity in Complex Lysates
Protein Assays to Guide Downstream Process Decisions
Scientists at Merck relied on Simple Western to quantify empty/full ratios of Coxsackievirus A21 (CVA21) by raising custom antibodies that target CVA21 capsid proteins. Simple Western protein size separation provided specificity of antibody reactivity to CVA21 capsid proteins. Adapted with permission (CC BY 4.0)1
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Isoforms, Fragments, and Aggregates
Therapeutic Antibodies
Simple Western protein size separation monitors therapeutic antibody production and aggregation directly in fed-batch and perfusion culture samples with only a single antibody. BisAb: bispecific antibody; HC: heavy chain. Adapted with permission (CC BY 4.0).2
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Degradation Products
Resolve and Quantify Full-Length Protein from Degradation Production
Immunoassay with protein size information using Simple Western is featured in a publication in Nature to measure full-length protein expression with simultaneous total protein detection (shown in blue) for accurate normalization of protein expression. Lower bands are likely degradation or incompletely synthesized products which are all detected by the same antibody and would not be resolved by ELISA or MSD assays. Adapted with permission (CC BY 4.0).4
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Oligomers and Post-Translational Modifications
Protein Biomarkers of Disease Pathogenesis
Teasing apart high molecular weight oligomers and deglycosylated isoforms of a biomarker implicated in multiple sclerosis, schizophrenia, and long COVID. Adapted with permission (CC BY 4.0).5
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How Simple Western Assays Can Help You
Meet Our Family of Products
Simple Western™ is a next-generation hands-free capillary immunoassay platform combining protein separation with sensitive chemiluminescence and fluorescence immunodetection using conventional primary and conjugated secondary antibodies.
Explore our Simple Western Instruments
Types of Protein Size Assays
Immunoassays
Simple Western immunoassays provide protein size separation information in addition to immunodetection of protein target(s) of interest. Simple Western merges the separative performance of capillary electrophoresis with the specificity of antibody-based detection.
Learn More about Immunoassays »
Total Protein Assays
In addition to immunoassay detection, Simple Western provides simultaneous total protein detection, which can be used to normalize protein expression data for accurate quantification of protein targets. For applications requiring normalizing protein signal, normalization to total protein is more reliable using a housekeeping protein as the normalizing denominator.
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Multiplex Assays
Get more out of your 3 µL sample! Multiplexing based on molecular weight gives you the flexibility you need to detect multiple protein targets that vary in molecular weight within the same capillary. With Jess, you have the ability to multiplex up to three channels to detect protein targets, and there is an extra channel for total protein normalization. Combine RePlex™ on Jess and Abby provides high-fidelity serial rounds of immunoprobing by removing the antibodies from the first probing cycle for a second probing cycle for immunoassay or total protein assay.
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Multi-Antigen Serology Assays
Size-based separation enables high-specificity multi-antigen serology testing for detailed insight into the humoral response beyond what ELISA can provide.
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P Gillespie, R Rustandi, A Swartz et al. (2023) Quantitation of coxsackievirus A21 viral proteins in mixtures of empty and full capsids using capillary western Hum Gene Ther 34:66-77. PMID: 36503264.
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A Aziz, N Majewska et al. (2020) Perfusion reduces bispecific antibody aggregation via mitigating mitochondrial dysfunction-induced glutathione oxidation and ER stress in CHO cells Sci Rep 10:16620. PMID: 33024175.
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B Howng, M Winter, C LePage et al. (2021) Novel ex vivo zymography approach for assessment of protease activity in tissues with activatable antibodies Pharmaceutics 13:1390. PMID: 34575469.
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S Albers, E Allen, N Bharti et al. (2023) Engineered tRNAs suppress nonsense mutations in cells and in vivo Nature 618:842-848. PMID: 37258671.
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B Charvet, J Pierquin, J Brunel et al. (2021) Human endogenous retrovirus type W envelope from multiple sclerosis demyelinating lesions shows unique solubility and antigenic characteristics Virol Sin 36:1006-1026. PMID: 33770381.