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Sample preparation for proteomics

ABSTRACT: Urea-containing buffer solutions are generally used in proteomic studies to aid protein denaturation and solubilisation during cell and tissue lysis. It is well-known, however, that urea can lead to the carbamylation of peptides and proteins and; subsequently, incomplete digestion of proteins. In our work, we sought for ways to retain urea in the digestion protocol to reduce sample handling, while minimizing urea-related drawbacks.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human) Rattus Rattus (black Rat)

TISSUE(S): Spleen, Melanocyte, Pancreas

SUBMITTER: Lazaro Hiram Betancourt Nunez

LAB HEAD: Gyorgy Marko-Varga

PROVIDER: PXD009426 | Pride | 2018-06-06

REPOSITORIES: Pride

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			Action	DRS
	20170108_LysC-Tryp_1A.raw	Raw
	20170108_LysC-Tryp_1B.raw	Raw
	20170108_LysC-Tryp_1C.raw	Raw
	20170108_LysC-Tryp_25C_1A.raw	Raw
	20170108_LysC-Tryp_25C_1B.raw	Raw

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Publications

Quantitative Assessment of Urea In-Solution Lys-C/Trypsin Digestions Reveals Superior Performance at Room Temperature over Traditional Proteolysis at 37 °C.

Betancourt Lazaro Hiram LH Sanchez Aniel A Pla Indira I Kuras Magdalena M Zhou Qimin Q Andersson Roland R Marko-Varga Gyorgy G

Journal of proteome research 20180607 7

Urea-containing buffer solutions are generally used in proteomic studies to aid protein denaturation and solubilization during cell and tissue lysis. It is well-known, however, that urea can lead to carbamylation of peptides and proteins and, subsequently, incomplete digestion of proteins. By the use of cells and tissues that had been lysed with urea, different solution digestion strategies were quantitatively assessed. In comparison with traditional proteolysis at 37 °C, urea in-solution digest ...[more]

PMID: 29812944

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