Protein Interaction Crosslinking for Mass Spectrometry

Thermo Scientific™ MS-grade crosslinkers are high-quality reagents that are available in multiple packaging options and sizes. We offer extensive technical expertise and support for various applications, as well as validation of these products in workflows using Thermo Scientific mass spectrometers.

  • High quality—products manufactured in ISO 9001–certified facilities
  • Convenience—products available in multiple package sizes, including “no-weigh” format
  • Wide variety—available with different linker lengths, MS-cleavability and deuterium isotope labels
  • Technical support—web resources and technical service to help ensure successful results

Choose the right crosslinker for your mass spec application

 DSG (disuccinimidyl glutarate)BS2G-d0DSSODSBUDSSBS3
CrosslinkerDSGBS2G-d0DSSO

DSBU

DSSBS3
Spacer arm7.7 Å7.7 Å10.3 Å

12.5  Å

11.4 Å11.4 Å
Water solubleNoYesNo

No

NoYes
MS cleavableNoNoYes

Yes

NoNo
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 BS2G-d4BS3-d4
CrosslinkerBS2G-d4BS3-d4
Spacer arm7.7 Å11.4 Å
Water solubleYesYes
MS cleavableNoNo
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Standard
 DSG (disuccinimidyl glutarate)BS2G-d0DSSODSBUDSSBS3
CrosslinkerDSGBS2G-d0DSSO

DSBU

DSSBS3
Spacer arm7.7 Å7.7 Å10.3 Å

12.5  Å

11.4 Å11.4 Å
Water solubleNoYesNo

No

NoYes
MS cleavableNoNoYes

Yes

NoNo
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Isotopically labeled
 BS2G-d4BS3-d4
CrosslinkerBS2G-d4BS3-d4
Spacer arm7.7 Å11.4 Å
Water solubleYesYes
MS cleavableNoNo
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Featured product data

Comparison of BSA crosslinking efficiency by SDS-PAGE

Comparison of BSA crosslinking efficiency by SDS-PAGE. Different crosslinkers were incubated with BSA at molar excess of crosslinker to protein (e.g., 20- 100-, or 500-fold). Crosslinking efficiency is shown by decreased mobility by SDS-PAGE and varied by crosslinker type, solubility, and concentration.

BSA crosslinked peptide

BSA crosslinked peptide spectra identified by MS2-MS3 method and XLinkX using DSSO crosslinker. XlinkX software uses unique fragment ion patterns of MS-cleavable crosslinkers (purple annotation) to detect and filter crosslinked peptides for a crosslinked database search.

Graph showing number of BSA crosslinked peptides

Graph showing number of BSA crosslinked peptides identified using different noncleavable (BS3, DSS) and cleavable (DSSO) crosslinkers for various MSn methods. Both BS3 and DSS had similar numbers of crosslinked peptides identified for collision-induced fragmentation (CID) and high-energy CID (HCD) methods, but DSSO identified fewer crosslinked peptides. Because DSSO is MS-cleavable, the MS2-MS3 method can be used to identify more crosslinks when a linear-peptide database search is used. All crosslinkers showed similar numbers of identified crosslinked peptides by electron-transfer and higher-energy collision dissociation (EThcD).

Resources

For Research Use Only. Not for use in diagnostic procedures.