The Simple Western is Here Name Date

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Western Blots are Tedious 3

Western Blots are Inconsistent 4 Day 1 Day 2

Meet Simon Gel-free, blot-free, hands-free Simple Western Assays 5

Simon Gives You More Walk away assay convenience Fully analyzed data Ease of Use Automated start to finish Protocol standardization Accurate quantitation Greater reproducibility Automation Higher Data Quality 6

Simple Westerns are Easy 7 Prepare samples Start Simon Results automatically

Get More Information 8 MCF7 Ladder Traditional Western BlotSimple Western

More Quantitative 9

More Reproducible 10 CV=5.2%CV=11.5%CV=12.0%CV=7.3%CV=9.7% egarevAyadsruhTyadsendeWyadseuTyadnoM NickNancyTimJessica Experiments(n=11replicates) TargetProteinReproducibility Signal

Global Standardization 11

How Simple Westerns Work Immobilize Separate Immunoprobe Quantitate Signal

Limits of Detection Western Blot Simple Western 2.0 mg/mL KDa Ladder 2.0 mg/mL Ladder

Wide Dynamic Range of Detection 14 Ladder3.0 mg/mL KDa

Multiplexing Capabilities 15 Courtesy of Senior Scientist pharmaceutical evaluator AKT2 Thioredoxin Ladder Rep 1 Rep 2 Rep 3 Rep 4

Comprehensive Pathway Validation 100 Targets Detected Insulin/ AKT NF  B DNA Damage Cell CycleApoptosis JAK/ STAT EGFR 16

Monitoring Biological Response Phospho-STAT5Total STAT5 - + MW - + MW 17 Courtesy of Senior Scientist pharmaceutical evaluator

Simple Westerns Offer More FeatureSimple WesternWestern Blot Quantitation Reproducibility Automation & Time Savings Sensitivity Throughput 18

Complete Solution InstrumentKitsSoftware 19

20 Powering Protein Research PurityImagingSignalingAggregationCharacterization

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Supplemental Slides 22

How the Simple Western Works 23

Simple Western Assay Specs DescriptionSpecification Samples per run12 Sample required3 µg Total run time3-5 hrs Sizing range kDa* Sizing accuracy± 20% across sizing range Sizing CV10% Resolution± 10% difference in molecular weight at 50% baseline (similar to resolution on a 4-20% gradient or 10% non-gradient SDS- PAGE gel) Dynamic range3 logs Relative Signal Quantitation CV < 20% across sizing range SensitivityLow ng *Assay specifications apply only to the stated sizing range. However, proteins up to 200 kDa can be detected.