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Suspension trapping (S-trap) is compatible with typical protein extraction buffers and detergents for bottom up proteomics

ABSTRACT: Analysis of cells and tissue by bottom up proteomics starts with lysis, followed by in-solution digestion. Lysis buffers commonly used include detergents and other reagents for efficient solubility of the proteins. However these reagents are, for the most part, incompatible with downstream analytical instrumentation. One method for in-solution digestion and cleanup, termed Suspension Trapping (S-Trap), has been recently introduced. We present evaluation of the compatibility of commonly used lysis buffers with S-trap: SDS, urea, NP-40, RIPA and SDS with DTT (SDT). We show that S-trap is compatible with all the tested buffers, with SDS and SDT performing the best. Based on this data, we anticipate that the method will transform experimental planning for mass spectrometry based proteomics, making it far more flexible and tolerable of various lysis buffers.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Yishai Levin

LAB HEAD: Yishai Levin

PROVIDER: PXD011665 | Pride | 2019-02-28

REPOSITORIES: Pride

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			Action	DRS
	HF2_7576_YLE_NP40-1_120718.raw	Raw
	HF2_7576_YLE_NP40-2_120718.raw	Raw
	HF2_7576_YLE_NP40-3_120718.raw	Raw
	HF2_7576_YLE_RIPA-1_120718.raw	Raw
	HF2_7576_YLE_RIPA-2_120718_180716020844.raw	Raw

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Publications

Suspension Trapping (S-Trap) Is Compatible with Typical Protein Extraction Buffers and Detergents for Bottom-Up Proteomics.

Elinger Dalia D Gabashvili Alexandra A Levin Yishai Y

Journal of proteome research 20190220 3

The analysis of cells and tissue by bottom-up proteomics starts with lysis, followed by in-solution digestion. Lysis buffers commonly used include detergents and other reagents for achieving efficient protein solubility. However, these reagents are, for the most part, incompatible with downstream analytical instrumentation. One method for in-solution digestion and cleanup, termed suspension trapping (S-Trap), has been recently introduced. We present an evaluation of the compatibility of commonly ...[more]

PMID: 30761899

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