Quickly remove detergents from protein samples to improve mass spectrometry results.
Thermo Scientific Pierce Detergent Removal Spin Columns efficiently remove high concentrations of detergents from 10µl to 1ml samples with minimal sample loss. The affinity resin effectively and specifically binds a wide variety of detergents and surfactants that are commonly used in protein extraction and biological sample preparation. The four available sizes of prepacked spin columns provide an especially convenient and rapid format for treating protein and peptide solutions to remove interfering detergents before downstream analysis by mass spectrometry and other techniques.
Highlights:
- Fast and efficienty protocol
- Removes >95% of detergents with high protein recovery
- Helps improves mass spectrometry peptide coverage
About Pierce Detergent Removal Resin:
Detergent removal from peptide samples is a challenge, especially for MS analysis in which even low detergent concentrations contaminate instruments and interfere with column binding, elution and ionization. Experiments demonstrate that Pierce Detergent Removal Resin effectively removes a variety of detergents from BSA and HeLa cell lysate tryptic digests, enabling improved LC-MS/MS and MALDI-MS analysis. After processing samples, the high baseline caused by detergents is reduced or eliminated. Analysis of the digested HeLa cell lysates by LC-MS/MS resulted in an approximate four-fold increase of identified peptides in the treated sample compared to the detergent-contaminated sample and equivalent numbers of peptides compared to a control sample, indicating minimal losses of peptides.
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| Protocol summary for Thermo Scientific Pierce Detergent Removal Spin Columns. |
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Peaks corresponding to detergents are eliminated in processed samples, allowing reliable peptide/protein identification. BSA tryptic digest (0.1ml, 100µg) containing a detergent was processed through 0.5ml of Pierce Detergent Removal Resin and subjected to MALDI-MS analysis on a MALDI-Orbitrap Mass Spectrometer. Similar results were produced for samples containing CHAPS, NP-40 and SDS (data not shown). |
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Effective detergent removal enables greater peptide identification. A tryptic digest of HeLa cell lysate (0.1ml, 100µg) containing 1% SDS was processed through 0.5ml 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. |
| Detergent are effectively removed with high protein recovery |
| Detergent |
Detergent Removal (%) |
BSA Recovery (%) |
| SDS, 2.5% |
99 |
95 |
| Sodium deoxycholate, 5% |
99 |
100 |
| CHAPS, 3% |
99 |
90 |
| Octyl glucoside, 5% |
99 |
90 |
| Octyl thioglucoside, 5% |
99 |
95 |
| Lauryl maltoside, 1% |
98 |
99 |
| Triton* X-100, 2% |
99 |
87 |
| Triton X-114, 2% |
95 |
100 |
| NP-40, 1% |
95 |
91 |
| Brij*-35, 1% |
99 |
97 |
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| Note: Samples (0.1ml containing 100µg BSA and detergent) were processed through 0.5ml of Pierce Detergent Removal Resin and the percent detergent removed was determined. Similar results were produced for insulin (5.7 kDa), α-lactalbumin (14.2 kDa) and carbonic anhydrase (29 kDa) (data not shown). |
Related Products:
Pierce Cell Lysis Reagents
Surfact-Amps Purifed Detergent Solutions
Pierce C-18 Spin Columns |
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