After isolation of peptides, salts and buffers can be removed using reversed phase (RP) resins, of which the C18 matrix is the most ideal for the capture of hydrophobic peptides.  The peptides bind to reverse-phase columns in high-aqueous mobile phase, salts and buffers are washed off, and the peptides are eluted using a high-organic mobile phase. However, hydophilic peptides, including phosphopeptides, may bind to C18 resins poorly, so graphite spin columns are recommended for better peptide recovery. However, C18 and graphite resins do not efficiently remove detergents, which can contaminate instruments and interfere with column binding, elution and ionization. The Thermo Scientific Detergent Removal products efficiently bind to and remove high concentrations of a wide variety of detergents that are commonly used during sample processing.

  • Comprehensive—resins and kits for multiple peptide clean-up strategies
  • Optimized—all products are designed to maximize peptide yield and LC-MS compatibility
  • Convenient—easy-to-use spin tip and columns enable more rapid clean-up of peptides
  • Flexible—devices can be used with either cultured mammalian cells or tissues
  • Validated—all products have been fully tested and processed samples have been analyzed using Thermo Scientific Mass Spectrometers
     

Choose the right peptide clean up products for your application

 
 Pierce Peptide Desalting columnsPierce C18 TipsPierce C18 Spin ColumnsPierce C18 Spin TipsGraphite Spin Columns
Primary applicationHigh capacity peptide desalting using microcentrifugePeptide desalting using micropipettePeptide desalting using microcentrifugeStage tip peptide desalting using microcentrifugePhosphopeptide desalting using microcentrifuge
FormatSpin columnTipSpin columnSpin tipSpin column
Maximum binding capacity5 mg10 µg or 80 µg30 µg10 µg100 µg
Maximum loading volume300 µL10 µL or 100 µL150 µL20 µL500 µL
Processing time15 min5 min25 min5-10 min10 min
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Featured data

Peptide recovery of native and TMT-labeled peptides

Figure 1. Peptide recovery of native peptides at various loads using the Pierce Peptide Desalting Spin Columns. Digested HeLa extracts (5–5,000 μg) were loaded onto Pierce Peptide Desalting Spin Columns and processed according to the protocol. Total recovered desalted peptides were estimated using the Pierce Quantitative Colorimetric Peptide Assay.

Peptide recovery of native and TMT-labeled peptides

Figure 2. Peptide recovery of native and TMT-labeled peptides. Pierce Peptide Desalting Spin Columns provide excellent binding and recovery characteristics for native and TMT-labeled peptide samples in preparation for MS. The columns effectively remove excess unreactive TMT label along with contaminating salts. Digested HeLa extract (500 µg) was labeled with TMT isobaric tags and loaded onto each column and processed according to the protocol. Total desalted peptide recovered was estimated using the Pierce Quantitative Colorimetric Peptide Assay.

Thermo Scientific™ Pierce™ C18 Spin Tips outperform other popular C18 tips.

Figure 3. Thermo Scientific Pierce C18 Spin Tips outperform other popular C18 tips. BSA tryptic digests were analyzed on an LTQ Orbitrap XL ETD Mass Spectrometer after processing 10 μg aliquots of the same digest with Pierce C18 Spin Tips or ZipTip™ Pipette Tips (10µL tips with 0.6 µL C18 resin). Base peak chromatograms of the peptide elution were extracted from the data sets to evaluate sample complexity and chromatographic resolution. MS results were analyzed using the SEQUEST search engine and the SwissProt database to determine protein sequence coverage.

Peptide recovery of native and TMT-labeled peptides

Figure 1. Peptide recovery of native peptides at various loads using the Pierce Peptide Desalting Spin Columns. Digested HeLa extracts (5–5,000 μg) were loaded onto Pierce Peptide Desalting Spin Columns and processed according to the protocol. Total recovered desalted peptides were estimated using the Pierce Quantitative Colorimetric Peptide Assay.

Peptide recovery of native and TMT-labeled peptides

Figure 2. Peptide recovery of native and TMT-labeled peptides. Pierce Peptide Desalting Spin Columns provide excellent binding and recovery characteristics for native and TMT-labeled peptide samples in preparation for MS. The columns effectively remove excess unreactive TMT label along with contaminating salts. Digested HeLa extract (500 µg) was labeled with TMT isobaric tags and loaded onto each column and processed according to the protocol. Total desalted peptide recovered was estimated using the Pierce Quantitative Colorimetric Peptide Assay.

Thermo Scientific™ Pierce™ C18 Spin Tips outperform other popular C18 tips.

Figure 3. Thermo Scientific Pierce C18 Spin Tips outperform other popular C18 tips. BSA tryptic digests were analyzed on an LTQ Orbitrap XL ETD Mass Spectrometer after processing 10 μg aliquots of the same digest with Pierce C18 Spin Tips or ZipTip™ Pipette Tips (10µL tips with 0.6 µL C18 resin). Base peak chromatograms of the peptide elution were extracted from the data sets to evaluate sample complexity and chromatographic resolution. MS results were analyzed using the SEQUEST search engine and the SwissProt database to determine protein sequence coverage.

Video of C18 spin tips in action


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