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SDS Removal with HILIC microSPE Columns SDS Removal with HILIC microSPE Column

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Peptide sequencing labs frequently use sodium dodecyl sulfate (SDS) to solubilize peptides and proteins from electrophoresis gels. Unfortunately, the SDS in the samples often ruins subsequent runs by reversed-phase HPLC (RPC). Moreover, in automated peptide sequencers, SDS in samples can lead to bubble formation in the sample intake lines, which blocks further sample intake. SDS can also accumulate on the PVDF membranes used for sequencing, washing off at an inconvenient time. All in all, it's a good idea to get it out of your samples.

Use of a HILIC HPLC Jeno's technique for SDS or other detergents, or by pre-treatment of the sample with a HILIC MiniSpin micro-SPE Tip (p/n: SUM HIL) can remove detergents and contaminating dyes prior to injection onto your RPC column.

In this example, 8.6% SDS was removed by HILIC from a peptide mixture prior to an RPC analysis with a HILIC microSPE tip from Glygen, Corp..

Data courtesy of Britt-Marie Olsson (Stockholm Univ.)

The size of the microSPE column is determined by the capacity needed for the retained molecules only, since the SDS comes off during the loading (in 60-80% ACN) and does not take up capacity on the HILIC cartridge. Peptide is eluted at 100% water 30mM ammonium formate and is thus ready for direct RPC loading, or for direct ESI-MS analysis.

See Spin columns for part numbers PUMSCHY1203 or PMASCHY1203 for individual Spin Columns or UltraMicroSpin or MicroSpin for packages of HILIC MiniSpin columns/Tip columns, SUM HIL, STP HIL and SEM HIL.

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Last Updated: 08/20/21