Detergent Removal from Peptides

The Thermo Scientific Pierce Detergent Removal Resins are provided in convenient spin-column or plate formats that quickly and efficiently remove ionic, nonionic, and/or zwitterionic detergents from protein or peptide samples to improve compatibility with downstream applications. Two formulations are available that are optimized to remove detergents from peptide samples with different concentration ranges. The Thermo Scientific HiPPR (high protein and peptide recovery) products are recommended for peptide samples ≤100 μg/mL. The standard Pierce Detergent Removal Resin products are ideal for peptide samples >100 μg/mL.

  • High performance—removes detergent with >90% recovery and no sample dilution
  • Versatile—effectively removes a wide variety of detergents from peptide or protein samples
  • Optimized—separate formulations for samples with peptide concentrations ≤ or >100 μg/mL
  • Flexible—available in various formats, including spin columns, 96-well spin plates, and loose resin
  • Convenient—simple method that helps to improve MS peptide coverage

Choose the right peptide detergent removal product for your application

  HiPPR Detergent Removal Spin Columns HiPPR Detergent Removal Spin Kit HiPPR Detergent Removal Spin Plates Detergent Removal Spin Columns (4 sizes) Detergent Removal Resin Detergent Removal Spin Plates
  HiPPR detergent removal column HiPPR detergent removal spin kit HiPPR detergent removal spin plates Detergent Removal Spin Columns Detergent Removal Resin Detergent Removal Spin Plates
Sample volume range(s) 100 µL 25– 200 µL 100 µL
  • 10– 25 µL
  • 25– 100 µL
  • 150– 500 µL
  • 500– 1,000 µL
10– 2,500 µL 20– 100 μL
Recommended peptide sample concentration range 1– 100 µg/mL 1–100 µg/mL 1–100 µg/mL >100 µg >100 µg >100 µg
Format Pre-filled spin columns 5 mL resin + 0.8 mL empty spin columns 96-well pre-filled spin plate Pre-filled spin columns 10 mL resin 96-well pre-filled spin plate
Resin bed volume(s) 0.1 mL 0.025–0.2 mL 0.1 mL
  • 0.125 mL
  • 0.5 mL
  • 2 mL
  • 4 mL
0.125 mL– 10 mL 0.55 mL
Cat. No. 88306 88305 88307 87777 87780 88304

Table 1. Results using standard Thermo Scientific Pierce Detergent Removal Spin Column, 0.5 mL.

Detergent removal efficiency and protein recovery. BSA sample (25–200 µL) + detergent in 0.15 M NaCl, 0.05% sodium azide was mixed with equal volume of detergent removal resin (2x volume for CHAPS removal).

Detergent Sample volume (µL) Protein quantity (µg) Detergent removal (%) Protein recovery (%)
SDS (1%) 25 0.375 >99 98
50 0.75 >99 97
100 1.5 >99 100
200 3.0 >99 100
Triton X-100 (1%) 25 0.375 >95 82
50 0.75 >95 86
100 1.5 >95 86
200 3.0 >95 93
NP-40 (0.75%) 25 0.375 95 90
50 0.75 96 94
100 1.5 97 91
200 3.0 97 97
CHAPS (1%) 25 0.375 95 64
50 0.75 97 70
100 1.5 98 78
200 3.0 98 75

Figure 1. Results using standard Pierce Detergent Removal Spin Column, 0.5 mL. Tryptic digests (0.1 mL, 100 μg) containing detergent were each processed through 0.5 mL of Pierce Detergent Removal Resin and subjected to LC-MS/MS analysis. Top row: Base peak LC-MS chromatograms. Bottom row: Integrated mass spectra. Similar results were produced for Brij-35 detergent, octyl glucoside, octyl thioglucoside, and SDS (data not shown).

Figure 2. Results using Pierce Detergent Removal Spin Columns, 0.5 mL. 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. Effective detergent removal enables greater peptide identification.

Table 2. Results using Thermo Scientific HiPPR Detergent Removal Resin.

Each column and plate well contained ~550 μL of detergent-removal resin slurry and 0.1 mL of sample.

Process format Detergent Detergent concentration (%) Detergent removal (%) BSA recovery (%)
0.5 mL Spin Column Sodium deoxycholate 5 99 100
Octyl glucoside 5 99 90
Octyl thioglucoside 5 99 95
Lauryl maltoside 1 98 99
Triton X-114 2 95 100
Brij-35 1 99 97
Tween 20 0.25 99 87
96-well Spin Plate SDS 5 99 89
Triton X-100 4 99 100
NP-40 1 95 100
CHAPS 5 99 100

 

Figure 3. Results using HiPPR Detergent Removal Resin. BSA (100 μg/mL) tryptic digests were prepared without detergent, in the presence of 0.5% Triton X-100 or spiked with 0.5% SDS following enzymatic digestion. Samples (0.1 mL) containing detergent were processed with the HiPPR Detergent Removal Resin and compared to unprocessed or detergent-free samples by LC-MS/MS. Results demonstrate that detergent removal is effective and produces results similar to those observed for samples containing no detergent.

Figure 4. Results using HiPPR Detergent Removal Resin. BSA (25 and 100 μg/mL) was digested in the presence and absence of detergents and the samples were processed for LC-MS/MS analysis. Effective detergent removal resulted in greater peptide identification and high MASCOT™ scores.

Table 1. Results using standard Thermo Scientific Pierce Detergent Removal Spin Column, 0.5 mL.

Detergent removal efficiency and protein recovery. BSA sample (25–200 µL) + detergent in 0.15 M NaCl, 0.05% sodium azide was mixed with equal volume of detergent removal resin (2x volume for CHAPS removal).

Detergent Sample volume (µL) Protein quantity (µg) Detergent removal (%) Protein recovery (%)
SDS (1%) 25 0.375 >99 98
50 0.75 >99 97
100 1.5 >99 100
200 3.0 >99 100
Triton X-100 (1%) 25 0.375 >95 82
50 0.75 >95 86
100 1.5 >95 86
200 3.0 >95 93
NP-40 (0.75%) 25 0.375 95 90
50 0.75 96 94
100 1.5 97 91
200 3.0 97 97
CHAPS (1%) 25 0.375 95 64
50 0.75 97 70
100 1.5 98 78
200 3.0 98 75

Figure 1. Results using standard Pierce Detergent Removal Spin Column, 0.5 mL. Tryptic digests (0.1 mL, 100 μg) containing detergent were each processed through 0.5 mL of Pierce Detergent Removal Resin and subjected to LC-MS/MS analysis. Top row: Base peak LC-MS chromatograms. Bottom row: Integrated mass spectra. Similar results were produced for Brij-35 detergent, octyl glucoside, octyl thioglucoside, and SDS (data not shown).

Figure 2. Results using Pierce Detergent Removal Spin Columns, 0.5 mL. 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. Effective detergent removal enables greater peptide identification.

Table 2. Results using Thermo Scientific HiPPR Detergent Removal Resin.

Each column and plate well contained ~550 μL of detergent-removal resin slurry and 0.1 mL of sample.

Process format Detergent Detergent concentration (%) Detergent removal (%) BSA recovery (%)
0.5 mL Spin Column Sodium deoxycholate 5 99 100
Octyl glucoside 5 99 90
Octyl thioglucoside 5 99 95
Lauryl maltoside 1 98 99
Triton X-114 2 95 100
Brij-35 1 99 97
Tween 20 0.25 99 87
96-well Spin Plate SDS 5 99 89
Triton X-100 4 99 100
NP-40 1 95 100
CHAPS 5 99 100

 

Figure 3. Results using HiPPR Detergent Removal Resin. BSA (100 μg/mL) tryptic digests were prepared without detergent, in the presence of 0.5% Triton X-100 or spiked with 0.5% SDS following enzymatic digestion. Samples (0.1 mL) containing detergent were processed with the HiPPR Detergent Removal Resin and compared to unprocessed or detergent-free samples by LC-MS/MS. Results demonstrate that detergent removal is effective and produces results similar to those observed for samples containing no detergent.

Figure 4. Results using HiPPR Detergent Removal Resin. BSA (25 and 100 μg/mL) was digested in the presence and absence of detergents and the samples were processed for LC-MS/MS analysis. Effective detergent removal resulted in greater peptide identification and high MASCOT™ scores.

Resources