Empore™ E4Technology
An enhanced E3Technology™ for low-cell and single-cell proteomics.
Key Features
Revolutionary features that transform proteomics sample preparation
Effective
Eliminate lysis step and surfactant usage keeping samples clean
Economical
To bridge gaps between genomics and proteomics products
Efficient
<15-min hands-on time
Robust
Zero technical barrier to even entry-level biomedical scientists
Versatile
E4tips™, E4filter™, E4cartridge™, and E4plate™ to satisfy different sample volumes, concentrations, quantities, and the need for automation
Stress-free
No liquid transfer, no concerns of any free-beads related issues (protein-bead ratios, beads sticking to tube walls and surfaces, cross-contaminations, etc.)
E4Technology™ Performance
Quantitative results demonstrating superior protein identification and reproducibility
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Application Notes
Figure 1. Qualitative assessment of E3technology (E3filter) for E. coli proteome analysis
(A-C) Comparison of the number of proteins, peptides, and PSMs between the E3filter, FASP, and SP4-GB approaches. Error bars represent three replicates.
(D-E) Overlapping analyses of proteins and peptides derived from the three methods
Figure 2. Qualitative assessment of E3technology (E3filter) for E. coli proteome analysis
(A-C) Comparison of the number of proteins, peptides, and PSMs between the E3filter, FASP, and SP4-GB approaches. Error bars represent three replicates.
(D-E) Overlapping analyses of proteins and peptides derived from the three methods
Figure 3. Qualitative assessment of E3technology (E3filter) for E. coli proteome analysis
(A-C) Comparison of the number of proteins, peptides, and PSMs between the E3filter, FASP, and SP4-GB approaches. Error bars represent three replicates.
(D-E) Overlapping analyses of proteins and peptides derived from the three methods
Revolutionary Proteomic Sample Preparation Technology
Empore™ E4Technology™ is designed to prepare LCMS-ready samples in an unprecedented manner.
The high efficiency and effectiveness along with versatility and low cost make it ideal for processing protein samples for basic research, clinical screening, and large-scale biomarker discovery. The “single vessel” nature of the technology also holds great value for “ultra-low-input” proteomics.
E4Technology™ Workflow
Save time and improve reproducibility with standardized, pre-packed tips
1
Cell treatment
Depending on the sample volume, add 2x pure methanol (100%) to the sample tube, and mix cells by gentle pipetting or vertexing.
2
Sample loading
Transfer cells to E4filters, centrifuge at 4,000 rpm for 1-2 min, and discard flow through.
3
Cell fixing
Add 200-500 µl methanol (90%) and incubate in ice or at 4°C for 0.5-2.0 hours.
4
Wash
Centrifuge at 4,000 rpm for 1-2 min. Discard flow through. Add 200-500 µl methanol (90%), centrifuge again, and discard flow through.
5
Digestion
Transfer E4filters to clean collection tubes, and add 50-200 µl 50 mM TEAB, desired enzyme (Trypsin or Trypsin/Lys-C mix) at a 1:50 ratio. Incubate at 37°C for 16-18 hours with gentle shaking.
6
Reduction and alkylation
Add 10 mM Tris (2-carboxyethyl) phosphine (TCEP) and 40 mM chloroacetamide (CAA) and incubate at 70°C for 30 min with gentle shaking.
7
Wash
Add acetic acid to the final concentration of 1%, centrifuge at 4,000 rpm for 1-2 min, and discard flow through. Add 200-500 µl 0.5% acetic acid in water, and centrifuge at 4,000 rpm for 1-2 min. Discard flow through.
8
Elution
Transfer E4filters to clean collection tubes. Do two sequential elution by adding 200-500 µl 60% acetonitrile/0.5% acetic acid in water (elution I), and 80% acetonitrile/0.5% acetic acid in water (elution II), centrifuge at 2,000-4,000 rpm for 1-2 min, pool the elution, dry, store at -80°C until LCMS analysis.
Products Selection Guide
Choose the right E4 procducts for your application
Resources for E4Technology™
Empore™ Proteomics catalog