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 ColumnsHiPPR Detergent Removal Spin KitHiPPR Detergent Removal Spin PlatesDetergent Removal Spin Columns (4 sizes) Detergent Removal ResinDetergent Removal Spin Plates
 HiPPR detergent removal columnHiPPR detergent removal spin kitHiPPR detergent removal spin platesDetergent Removal Spin ColumnsDetergent Removal ResinDetergent Removal Spin Plates
Sample volume range(s)100 µL25– 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 range1– 100 µg/mL 1–100 µg/mL1–100 µg/mL>100 µg>100 µg>100 µg
FormatPre-filled spin columns5 mL resin + 0.8 mL empty spin columns96-well pre-filled spin platePre-filled spin columns10 mL resin96-well pre-filled spin plate
Resin bed volume(s)0.1 mL0.025–0.2 mL0.1 mL
  • 0.125 mL
  • 0.5 mL
  • 2 mL
  • 4 mL
0.125 mL– 10 mL 0.55 mL
Cat. No.883068830588307877778778088304

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).

DetergentSample volume (µL)Protein quantity (µg)Detergent removal (%)Protein recovery (%)
SDS (1%)250.375>9998
500.75>9997
1001.5>99100
2003.0>99100
Triton X-100 (1%)250.375>9582
500.75>9586
1001.5>9586
2003.0>9593
NP-40 (0.75%)250.3759590
500.759694
1001.59791
2003.09797
CHAPS (1%)250.3759564
500.759770
1001.59878
2003.09875

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 formatDetergentDetergent concentration (%)Detergent removal (%)BSA recovery (%)
0.5 mL Spin ColumnSodium deoxycholate599100
Octyl glucoside59990
Octyl thioglucoside59995
Lauryl maltoside19899
Triton X-114295100
Brij-3519997
Tween 200.259987
96-well Spin PlateSDS59989
Triton X-100499100
NP-40195100
CHAPS599100

 

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.

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).

DetergentSample volume (µL)Protein quantity (µg)Detergent removal (%)Protein recovery (%)
SDS (1%)250.375>9998
500.75>9997
1001.5>99100
2003.0>99100
Triton X-100 (1%)250.375>9582
500.75>9586
1001.5>9586
2003.0>9593
NP-40 (0.75%)250.3759590
500.759694
1001.59791
2003.09797
CHAPS (1%)250.3759564
500.759770
1001.59878
2003.09875

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 formatDetergentDetergent concentration (%)Detergent removal (%)BSA recovery (%)
0.5 mL Spin ColumnSodium deoxycholate599100
Octyl glucoside59990
Octyl thioglucoside59995
Lauryl maltoside19899
Triton X-114295100
Brij-3519997
Tween 200.259987
96-well Spin PlateSDS59989
Triton X-100499100
NP-40195100
CHAPS599100

 

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.