ProteinSimple Automated Western Blot: Reinventing Western Blotting


Reinventing Western Blotting Through Automation

Many scientists have experienced the challenges plaguing the traditional Western Blot test. BioAgilytix provides an innovative alternative through its ProteinSimple Peggy Sue platform, which automates Western blot assays to overcome the obstacles related to reproducibility, sensitivity, quantifiability, and speed seen in traditional Western blotting.

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proteinsimple peggy sue platform analytical device

How the ProteinSimple Platform Works

ProteinSimple is an automated capillary immunoblotting system that enables the development of assays when there is only one antibody available, and the target analyte requires separation based on size and/or charge prior to analysis. ProteinSimple Peggy Sue is able to separate proteins in two ways – by size and by charge – via ProteinSimple’s proprietary, photoactivated capture chemistry, yielding a quantifiable chemiluminescent signal. This can all be done using as little as 5 μL of sample.

When dealing with size-based assays, the ProteinSimple platform automatically performs sample loading, size-based protein separation, immunoprobing or total protein labeling, washing, detection and data analysis. As described on the ProteinSimple website, up to 96 samples, including controls and molecular weight ladders, can be processed at once, in as little as 24 hours.

When processing charge-based assays, ProteinSimple Peggy Sue automates sample loading, charge-based protein separation, immunoprobing, detection, and data analysis. It can similarly process up to 96 samples in less than 24 hours and allows for analysis of small isoelectric point differences of target proteins. Like the size-based assays, these charge-based assays take place in a capillary, where they are automatically separated by charge and resolve according to expected pl values. In either separation scenario, automation mitigates manual variability and cuts out transfer of proteins to yield consistent results.

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bioagilytix platform comparison guide

What Platform is Best for Your Project?

Use our Platform Comparison Guide to evaluate the optimal method and technology for your assay, based on its unique characteristics and performance requirements.

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Ideal ProteinSimple Applications: For Biomarkers in a Wide Range of Matrices

ProteinSimple has essentially reinvented Westerns, removing the issues and frustrations with reproducibility, sensitivity, quantifiability, and speed seen in traditional methods. It offers a reliable, rapid alternative to traditional Western blot analysis for protein separation, identification, and quantitation. ProteinSimple Peggy Sue provides the added flexibility of quantitating biological responses from two different perspectives. If these parameters are critical to your project, ProteinSimple may be the right platform choice.

At BioAgilytix, ProteinSimple is most often used by our scientists in work related to biomarkers and a wide variety of matrices like plasma / serum, tissue homogenates, yeast and bacteria extracts, PBMCs and other mammalian cell lysates, and various plant tissues.

Key ProteinSimple Benefits: Improved Sensitivity and Linear Dynamic Range

ProteinSimple assays combine capillary electrophoresis, a traditional separation technique, with an immunoassay, a well-known detection technique, allowing BioAgilytix to perform quantitative Westerns with sensitivity and reproducibility unmatched by traditional Western blot methods.

Through automation, ProteinSimple mitigates the variability seen in manual processes that used to impact Western blotting protocols. The blotting step is removed, so inconsistencies in protein transfer are eliminated, dramatically improving time to result and overall data reliability. Additionally, these Simple Western assays have a linear dynamic range, so that proteins can be detected accurately with high throughput.

proteinsimple analysis report

Top Benefits of ProteinSimple:

  • High Throughput
  • Low Sample Volume (5 μL)
  • Improved Sensitivity Over Traditional Westerns
  • High Precision
  • Linear Dynamic Range
  • Robust & Rugged
  • Quantitative & Qualitative Assays
  • Separation Based on Size and/or Charge (IEF)
  • Total Protein Analysis
proteinsimple automation paper

Platform In Action:

Ligand Binding Assays in the 21st Century Laboratory: Automation

Learn how automation is helping analysts address growing pressures to increase efficiency and output with limited resources.

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Bioagilytix’s ProteinSimple Platform Expertise

BioAgilytix offers ProteinSimple Peggy Sue as part of an extensive portfolio of platforms. Our scientists have used this technology for a number of studies related to biomarker assessment in a variety of matrices including plasma/serum, tissue homogenates, PBMCs and other mammalian cell lysates, and various plant tissues.

Speak to our scientists today to learn how we will help support your Western blotting protocols with accurate, rapid results.

scientist performing proteinsimple assays

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