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

Standard
Isotopically labeled


Crosslinker	DSG	BS2G-d0	DSSO	DSBU	DSS	BS3
Spacer arm	7.7 Å	7.7 Å	10.3 Å	12.5 Å	11.4 Å	11.4 Å
Water soluble	No	Yes	No	No	No	Yes
MS cleavable	No	No	Yes	Yes	No	No
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Crosslinker	BS2G-d4	BS3-d4
Spacer arm	7.7 Å	11.4 Å
Water soluble	Yes	Yes
MS cleavable	No	No
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Standard


Crosslinker	DSG	BS2G-d0	DSSO	DSBU	DSS	BS3
Spacer arm	7.7 Å	7.7 Å	10.3 Å	12.5 Å	11.4 Å	11.4 Å
Water soluble	No	Yes	No	No	No	Yes
MS cleavable	No	No	Yes	Yes	No	No
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Isotopically labeled


Crosslinker	BS2G-d4	BS3-d4
Spacer arm	7.7 Å	11.4 Å
Water soluble	Yes	Yes
MS cleavable	No	No
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Featured product data

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 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 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.