PXD003653

PXD003653 is an original dataset announced via ProteomeXchange.

Title	Protein precipitation of diluted samples with acetone leads to higher protein recovery and reproducibility in comparison with TCA/acetone approach
Description	Proteomic approaches are extremely valuable in many fields of research, where mass spectrometry methods have gained an increasing interest especially due to the ability to perform quantitative analysis. Nonetheless, sample preparation prior to mass spectrometry analysis is of the utmost importance. In this work two protein precipitation approaches, widely used for cleaning and concentrating protein samples, were tested and compared in a case of very diluted samples solubilized in a strong buffer (containing SDS). The amount of protein recovered after acetone and TCA/acetone precipitation was assessed, as well as the protein identification and relative quantification by SWATH-MS yields which were compared with the results from the same sample without precipitation. With this study it was possible to conclude that, in the case of diluted samples in denaturing buffers, the use of cold acetone as precipitation protocol is more favourable than the use of TCA/acetone in terms of reproducibility in protein recovery, number of identified and quantified proteins. Furthermore, the reproducibility in relative quantification of the proteins is even higher in samples precipitated with acetone when comparing to the original sample.
HostingRepository	PRIDE
AnnounceDate	2016-04-25
AnnouncementXML	Submission_2016-04-25_03:30:14.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Cátia Santa
SpeciesList	scientific name: Rattus norvegicus (Rat); NCBI TaxID: 10116;
ModificationList	S-carboxamidoethyl-L-cysteine
Instrument	TripleTOF 5600

Revision	Datetime	Status	ChangeLog Entry
0	2016-02-17 01:58:05	ID requested
⏵ 1	2016-04-25 03:30:15	announced

Santa C, Anjo SI, Manadas B, Protein precipitation of diluted samples in SDS-containing buffer with acetone leads to higher protein recovery and reproducibility in comparison with TCA/acetone approach. Proteomics, 16(13):1847-51(2016) [pubmed]

curator keyword: Technical

submitter keyword: Protein Precipitation

Acetone

TCA/acetone

SWATH

Bruno Manadas
contact affiliation	Center for Neurosciences and cell biology, Coimbra, Portugal
contact email	bmanadas@gmail.com
lab head
Cátia Santa
contact affiliation	Center for Neuroscience and Cell Biology
contact email	catiajmsanta@gmail.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD003653
     1. Label: PRIDE project
     2. Name: Protein precipitation of diluted samples with acetone leads to higher protein recovery and reproducibility in comparison with TCA/acetone approach