Smile! Peptide fractionation simplified

Simple, fast and reproducible peptide fractionation giving increased proteomic depth

The challenge:

Preparing samples with complex proteomes e.g. human cell lines or tissues, or samples with a wide dynamic range in protein abundances for in depth proteomic characterizations is a compromise between effort, cost and method complexity.

The solution:

The PreOmics® iST-Fractionation Add-on is based on dipole-moment and mixed-phase interactions (patent pending). Samples are prepared using the PreOmics® iST kits until the WASH step and then fractionated using PreOmics® iST-Fractionation Add-on kit. The fractionation buffers are prepared ready for use and the method takes just 10 minutes. The resulting fractions provide a significant increase in the peptide and protein identifications from a single sample.

Apply! Simple, streamlined workflows

1. Samples

Follow iST or iST-NHS protocol until WASH steps.

2. Fraction-1

Swap to a collection container. Add FRACTION-1 to the cartridge and spin.

3. Fraction-2

Change to a new collection container. Add FRACTION-2 to the cartridge and spin.

4. Fraction-3

Change to a new collection container. Add FRACTION-3 to the cartridge and spin.

5. MS analysis

Dry the samples and resuspend them in LC-LOAD. Inject and analyze to see improved peptide and protein identifications.

Results

**Figure 1:** Using 6x10⁵ HeLa cells as starting sample. The left graph shows protein identifications for the three fractions each vs total proteins found for the three fractions combined vs the amount of protein identifications for an unfractionated sample, similarly the right graph shows the same information for the peptide identifications.

**Figure 2:** Using human plasma (2 µL) as starting sample. The left graph shows protein identifications for the three fractions each vs total proteins found for the three fractions combined vs the amount of protein identifications for an unfractionated sample, similarly the right graph shows the same information for the peptide identifications.

PreOmics^®'

iST-Fractionation Add-on

values

Smile

Improved sample quality

Achieve proficient peptide fractionation easily
Working range: 1-100 µg
Typically 40-50% more protein identifications compared to an unfractionated sample*

* Depending on sample type and MS instrumentation

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