C18 Columns & Peptide Clean Up for Mass Spectrometry
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™ C18 Spin Tips||Pierce™ C18 Tips||Pierce™ C18 Spin Columns||Pierce™ Detergent Removal Spin Columns||Pierce™ HiPPR Detergent Removal Spin Columns|
|Primary Application||Concentrate, desalt peptides||Concentrate, desalt peptides||Concentrate, desalt peptides||Remove detergents||Remove detergents|
|Format||Spin tip||Tip||Spin column||Spin column||Spin column|
|Resin bed volume||20 µL||
|8 mg||.125-4 mL slurry||0.25-100 µL slurry|
|Binding capacity or loading volume||10 µg||80 µg||30 µg||0.1-1 mL
µL (1-100 µg/mL)
|Processing time||5-10 min||5 min||25 min||15 min||15 min|
|Order now||Order now||Order now||Order now||Order now|
Need to clean up phosphopeptides?
Concentrate and desalt phosphopeptides rapidly and efficiently using Thermo Scientific™ Graphite Spin Columns. These ready to use spin columns can clean up to 100 μg of phosphopeptides in less than 10 minutes.
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.
Improved recovery of representative hydrophilic phosphopeptides using graphite spin columns. Stable isotope-labeled A3 and B9 peptides (10 pmol) were acidified with 1% trifluoroacetic acid, processed according to instructions for C18 tips or the Pierce Graphite Spin Columns, and eluted with 50% acetonitrile/0.1% formic acid. The corresponding heavy isotope-labeled peptides (5 pmol) were spiked in the eluate, dried and resuspended in 0.1% formic acid. Samples were analyzed by targeted LC-MS/MS with the Orbitrap XL Mass Spectrometer to quantitate percent recovery. Peptides: A3= RPRAApTFPFR†, B9 = RTPKDpSPGIPPFR†. Position of heavy isotope labeled amino acid used for absolute MS quantitation.
Effective detergent removal enables greater peptide identification. A tryptic digest of HeLa cell lysate (0.1 mL, 100 µg) containing 1% SDS was processed through 0.5 mL of Pierce Detergent Removal Resin and subjected to LC-MS/MS analysis. The processed sample allowed similar numbers of identified peptides as digests containing no SDS. Peptide identification is greatly reduced in sample containing SDS.
Video of C18 spin tips in action
Other products for peptide clean up and detergent removal for mass spec
For Research Use Only. Not for use in diagnostic procedures.