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Simple Western systems

Gel-free, blot-free, hands-free

OVERVIEW

Simple Western™ platforms are the only systems on the market that fully automate traditional western blotting, providing data on 24 samples in only 3 hours or 96 samples overnight. Separate and analyze proteins by size from 2-440 kDa either by immunoassay or total protein analysis. There's even an automated charge-based assay utilizing isoelectric focusing if you're doing post-translational modification characterization. You'll get quantitative results, reproducibility that's spot on, and use less sample in the process. So, what's the big difference between Simple Western automated western blot systems and traditional western blot tests? Our Simple Western instruments are the only gel-free, blot-free, and hands-free capillary-based immunoassay platforms that integrate and automate the entire protein separation and detection process. So you'll have more time to get down to real science.

Bringing Traditional Western Blotting Into the 21st Century

Historically, the traditional Western blot has been widely used in the study of protein biochemistry, but it is notoriously labor-intensive, and limited in reproducibility and quantification, among other challenges. Developed in the 1970s, the original Western blotting technique has remained virtually the same with limited improvements made to meet the demands of researchers.

Simple Western completely eliminates the manual and time-consuming steps of traditional western blotting workflows including the long and variable transfer step and the manual incubation, washing and detection steps that lead to the high variability and poor reproducibility that has plagued traditional Western blotting workflows for years.

Simple Western's hands-free automation, rapid time to results, quantitation, high sensitivity and reproducibility make it the technology of choice to meet researcher’s Western blot analysis needs in the 21st Century.

Simple Western Automates the Manual and Time-consuming Steps of Traditional Western Blotting Workflows

What Is the Difference Between Simple Western Automated Western Blot Systems and Traditional Western Blot Workflows?

Simple Western platforms redefine your western blotting workflow by fully automating all assay steps including separation, immunodetection and data analysis. Simple Western platforms are based on capillary electrophoresis and Simple Western assays are performed in capillaries where SDS/heat-denatured proteins are separated by apparent molecular weight, immobilized covalently via a UV sensitive coating in the capillary, detected directly in the capillary by primary and secondary antibodies and analyzed by the integrated Compass Software.

By integrating and automating these previously manual workflow steps, Simple Western platforms overcome many of the limitations associated with traditional Western blotting workflows, significantly reducing hands-on time, and providing excellent reproducibility and assay transferability. Simple Western assays are quantitative, enabling generation of full dose response curves, and also include built-in total protein normalization so you can easily and accurately normalize your immunoassay data.

Simple Western is easy to use and also offers higher throughput and faster time to results. With only 30 min set-up times and walk away automation, you can get fully analyzed western blotting data on 25 samples in less than 3 hours on Jess, Abby, and Wes. If you want even higher throughput, Sally Sue or Peggy Sue can analyze up to 96 samples in a single run!

With all these advantages, Simple Western is rapidly becoming accepted as the preferred replacement for traditional Western blot protocols.

 
Simple Western System Jess Abby Wes Sally Sue Peggy Sue Traditional Westerns
Maximum samples per run 25 25 25 96 96 25
Run time for mass samples (size assays) <3 hours <3 hours <3 hours <14 - 19 hours <11 - 19 hours 2 days
Hands-on time 30 minutes 30 minutes 30 minutes 1 hour 1 hour 3.25 hours

Simple Western – Your Partner for Publication Perfect Data!

Increasingly, the scientific community is setting new and more rigorous standards for data reproducibility, transparency and rigor in grant submission guidelines and journal publications. With over a thousand publications and counting, Simple Western systems overcome the challenges associated with traditional western blotting workflows to produce publication-ready data that meet the highest standards of data integrity, reproducibility and rigor.

Automated Simple Western Platforms Provide Reproducible, Publication-ready Data

Automated Simple Western Platforms Provide Reproducible, Publication-ready Data

Key Benefits of Using Simple Western for Publication-ready Data:

  • Automated Simple Western platforms provide reproducible data, making it easy for other labs to reproduce & verify published results.
  • Generate complete electronic data files which include publication-quality images and quantitative results as well as all assay conditions, making it easy to upload to data repositories to meet data sharing requirements.
  • Perform total protein normalization in the same capillary as your immunoassay for more accurate results so you don’t have to rely on variable housekeeping proteins.
  • Perform reliable sequential immunoassays in the same capillary with RePlex™ rather than struggling with variable stripping & re-probing protocols.
  • Optimize your assay within hours making use of the vast selection of Simple Western certified antibodies.
  • Reliably detect low abundance proteins using highly sensitive chemiluminescence or fluorescence detection modes.
  • Get multiplexed western blotting data in the same sample using Jess’ fluorescence capabilities with IR, NIR and chemiluminescence channels.

Learn more  about how Simple Western systems empower researchers to publish faster while meeting journal requirements for rigor, reproducibility and data transparency.

MEET YOUR WESTERN PROTEIN ANALYSIS PROBLEM SOLVERS!

 Simple Western Jess Instrument: Automated Western Blotting System with Chemiluminescence, Fluorescence and RePlex

Jess

Size assays on up to 25 samples
Chemiluminescence and Fluorescence
Includes RePlex

Simple Western Abby Instrument: Automated Western Blotting System with Chemiluminescence and RePlex Features.

Abby

Size assays on up to 25 samples
Chemiluminescence
Includes RePlex

Simple Western Wes Instrument: Automated Western Blotting System with Chemiluminescence

Wes

Size assays on up to 25 samples
Chemiluminescence

Simple Western Sally Sue Instrument: Size Assay Based Protein Separation and Analysis System with Chemiluminescence

Sally Sue

Size assays on up to 96 samples
Chemiluminescence

Simple Western Peggy Sue Instrument: Size and Charge Assay Based Protein Separation, Identification and Quantitation System with Chemiluminescence

Peggy Sue

Size and charge assays on up to 96 samples
Chemiluminescence

Simple Western NanoPro 1000 Instrument: Charge Assay Based Protein Characterization and Analysis System with Chemiluminescence

NanoPro 1000

Charge assays on up to 96 samples
Chemiluminescence

Simple Western Assay: How does it work?

Two ways to get separation, identification and quantitation

You can separate your proteins two ways with Simple Western™ assays—by size or by charge. We've got systems that'll do one or the other, or both! No matter what option you choose, you'll get the separation you need, identification of your target protein, and truly quantitative data that lets you make the most accurate experimental decisions. Learn more.

Simple Western size-based western blotting assays

  • A Simple Western by size is an automated Western—no gels, no transfer devices, no blots, no film and no manual analysis. Just load your samples in Jess, Abby, Wes, Peggy Sue, or Sally Sue and press start—it's a complete, walk-away solution for protein separation and detection.
  • Jess, Abby, Wes, Peggy Sue, and Sally Sue automate all of the steps of traditional western blots by automatically including sample loading, size-based protein separation, immunoprobing or total protein labeling, washing, detection and data analysis.
  • Western blotting automation eliminates variability inherent to manual processes which limits reproducibility, while adding quantitation, and improving on time-to-result and overall data reliability.
  • The transition from traditional western blotting methods to automated western blots on Simple Western is simple—the same antibodies used for western blots can be used with our automated Simple Western assays.
  • Up to 96 samples can be processed at once, and Simple Western assays take just 3-19 hours to complete with sizing and quantitation results.
  • Separate and detect proteins as small as 2 kDa or as large as 440 kDa in only 3 hours.

How do Simple Western size-based assays work?

There are two ways to run Simple Western size-based assays—immunoassay or total protein. Just load your samples, start the instrument, and off to the races you go!

Simple Western immunoassays take place in a capillary. Samples and reagents are loaded into an assay plate and placed in Jess, Abby, Wes, Peggy Sue, or Sally Sue. As little as 40 nL of sample is loaded into the capillary automatically and proteins are separated by size as they migrate through stacking and separation matrices. The separated proteins are then immobilized to the capillary wall via a proprietary, photoactivated capture chemistry. Target proteins are identified using a primary antibody and immunoprobed using an HRP-conjugated secondary antibody and chemiluminescent substrate. The resulting chemiluminescent signal is detected and quantified.

Immunoassay Workflow Using Automated Western Blotting with Simple Western Technology

Interested in looking at all the proteins in your lysate? Just leave out the antibody and let Jess, Abby, Wes, Peggy Sue or Sally Sue do the work for you to add western blotting results of total protein in your sample. Target proteins separated by size are labeled with a biotin reagent and are detected by chemiluminescence using Streptavidin-HRP. At the end of the run, you've got relative quantitation for your protein of interest and total protein measurements of the sample.

Total Protein Assay Workflow Using Automated Western Blotting with Simple Western Technology

What does Simple Western size data look like? Simple Western size-based assay data is processed automatically for you in Compass software. Sample data is displayed by lane in a virtual-blot like image similar to traditional western blot results with one big exception—not only do you get more information, you get it as soon as the assay is complete. Quantitative results such as molecular weight, signal intensity (area), % area, and signal-to-noise for each immunodetected protein are presented in the results table automatically.

Lane View Data For Simple Western Size-based Assay

Just need total protein? We've got that covered too!

Automated western data for total protein detection

Total Protein detection. Decreasing concentrations of DNAK in Hela lysate and negative controls (15, 7.5 and 3.75 µg/mL).

If you're more familiar with capillary electrophoresis, you can see your results in a more traditional electropherogram view too.

Size based Simple Western electropherogram

Looking at big proteins? Try this on for size. Simple Western now has a wider molecular weight range capability, so you can detect proteins as large as 440 kDa.

Lane View of Automated Western Results From High Molecular Weight Targets

Detection of 3 high molecular weight targets. HECTH9, Talin and UBR5 were detected in K562 lysate using the 66-440 kDa Master Kit.

Reproducibility and quantitation

Simple Western assays are fully automated by Jess, Abby, Wes, Peggy Sue, or Sally Sue and easily standardized, making your results much more reproducible. And there\’s no blotting step, so inconsistencies caused by protein transfer are eliminated, which really improves your quantitation too!

PI3K expression Results Shown in Lane View From Automated Simple Western Application
PI3K table

Phosphoinositide 3-kinase (PI3K) expression in an MCF-7 lysate. Ten replicates (n=10) of MCF7 lysate at 1 µg/µL were analyzed using a PI3K-specific primary antibody. The predicted molecular weight of PI3K is 110 kDa. Simple Western assay data reported an average molecular weight of 107 kDa with a 0.5% CV for sizing and an 8.7% CV for signal intensity (area).

Dynamic range

Simple Western assays have a linear dynamic range of approximately 3 orders of magnitude, letting you detect proteins over a wider concentration range, not to mention with more accurate quantitation.

Quantitation of GSK-3 alpha with Simple Western

Dynamic range of detection for GSK-3 alpha. Lysate from K562 cells lysed in Bicine/CHAPs buffer was serially diluted and analyzed using a GSK-3 alpha antibody. The calculated dynamic range was 3.3 logs with an R² value of 0.9995.

Simple Western charge-based assays

A Simple Western charge assay is an automated assay just like a Simple Western size assay—without the use of gels, transfer devices, blots, film or manual analysis—but the data you get is very different. Because proteins are separated by the isoelectric point (pI), you'll be able to see extremely small isoelectric point differences. It's a new way of looking at discrete changes in very small samples that you might just be surprised by!

  • Peggy Sue and NanoPro 1000 automate traditional 1D isoelectric focusing slab gel protocols including sample loading, charge-based protein separation, immunoprobing, detection and data analysis.
  • Low abundance proteins in limited cell populations are a specialty and as few as 25 cells per assay are required.
  • Up to 96 samples can be processed at once and assays take just 19 hours to complete with reported pIs and quantitation results.
  • You can characterize actions of kinase inhibitor drugs or study on coprotein control mechanisms in primary tissues or fine needle aspirates.

How do Simple Western charge-based assays work?

Simple Western charge-based assays take place in a capillary. Just load your samples and off to the races you go!

Samples and reagents are loaded into an assay plate and placed in Peggy Sue or NanoPro 1000. Proteins and ampholytes are loaded into the capillary automatically and separated by charge and resolve according to their expected pI values. The separated proteins are then immobilized to the capillary wall via a proprietary, photoactivated capture chemistry. Target proteins are identified using a primary antibody and immunoprobed using an HRP-conjugated secondary antibody and chemiluminescent substrate. The resulting chemiluminescent signal is detected and quantitated.

Simple Western charge-based immunoassay

What Does Simple Western Charge Data Look Like? Simple Western charge-based assay data is processed automatically in Compass software. Sample data is displayed by electropherogram where it's easier to see minute changes. Quantitative results such as pI value, signal intensity (area), % area, and signal-to-noise for each immunodetected protein are presented in the results table automatically.

Charge-based assay data from Simple Western

Simple Western systems comparison chart

Which system is right for you?

Simple Western System Jess Abby Wes Sally Sue Peggy Sue NanoPro 1000
Simple Western size assays  
Simple Western charge assays        
Chemiluminescence
Fluorescence          
Replex™        
Maximum samples per run 25 25 25 96 96 96
Run time for max samples (size) < 3 hours < 3 hours < 3 hours 14-19 hours 11-19 hours N/A
Run time for max samples (charge) N/A N/A N/A N/A 11 hours 11 hours
Sample cooling (size) N/A N/A N/A 10°C 10°C N/A
Sample cooling (charge) N/A N/A N/A N/A 3°C 3°C
Operating humidity range 20-60% relative, non-condensing
Operating temp range 18-24°C
Power 100-230 V AC, 50/60 Hz
Dimensions (H x W x D) 33 x 33 x 52 cm 33 x 33 x 52 cm 33 x 33 x 52 cm 84 x 94 x 61 cm 84 x 94 x 61 cm 84 x 94 x 61 cm
Part number 004-650 004-680 004-600 004-700 004-800 004-109

 

  Immunoassay Size Specification Immunoassay Charge Specification
Description Chemiluminescence Fluorescence  
Sample required 0.3-1.2 µg lysate 0.6-1.2 µg 0.6-1.2 µg
Size or pI range 2-40, 12-230 and 66-440 kDa 2-40, 12-230 and 66-440 kDa Widest gradient ranges
from pI 3 to pI 10
Sizing CV <10% <10% <10%
Intra-assay CV <15% <15% <20%
Inter-assay CV <20%* <20%* <20%**
Resolution
(± percent difference in MW)
± 15-20% for MW
< 20 kDa
± 10% for MW
> 20 kDa
± 15-20% for MW
< 20 kDa
± 10% for MW
> 20 kDa
± 0.1 pI units
Quantitation CV <20% <20% <20%
Dynamic range up to 4 logs*** 3-4 logs 3 logs
Sensitivity Low pg High pg Low pg
Capillary 5 cm, 100 µm, 400 nL

*inter-assay CV is with system control
**percent peak area
***Jess/Wes with HDR detection mode.

Simple Western Applications

Cancer &
Immuno-Oncology

Simple Western Instrument Cancer Research Applications

 
 
SARS-CoV-2

Simple Western Instrument SARS-CoV-2 Research Applications

 
Cell & Gene Therapy

Simple Western Instrument Cell & Gene Therapy Research Applications

 
C&GT Western Blotting Solutions

Cell & Gene Therapy Western Blotting Solutions

 

Targeted Protein Degradation

Targeted Degradation

SARS-CoV-2 Serology

SARS-CoV-2 Multi-Antigen Serology Module

 
Exosome

Exosome

Vaccine Development

Vaccines Development

 
Neuroscience

Neuroscience