Repository logo
 

Assessment, Development, and Improvement of SDS Depletion Strategies in Mass Spectrometry Proteomic Workflows

dc.contributor.authorKachuk, Carolyn
dc.contributor.copyright-releaseYesen_US
dc.contributor.degreeMaster of Scienceen_US
dc.contributor.departmentDepartment of Chemistryen_US
dc.contributor.ethics-approvalNot Applicableen_US
dc.contributor.external-examinern/aen_US
dc.contributor.graduate-coordinatorMark Stradiottoen_US
dc.contributor.manuscriptsYesen_US
dc.contributor.thesis-readerHeather Andreasen_US
dc.contributor.thesis-readerPeter Wentzellen_US
dc.contributor.thesis-supervisorAlan Doucetteen_US
dc.date.accessioned2016-08-22T13:42:00Z
dc.date.available2016-08-22T13:42:00Z
dc.date.defence2016-08-10
dc.date.issued2016-08-22T13:42:00Z
dc.description.abstractIn proteomics, sodium dodecyl sulfate negatively affects trypsin digestion, chromatography, and mass spectrometry, thus, its effective depletion is critical to any protein sample workflow. A quantitative assessment of SDS depletion protocols reveals that acetone precipitation outperforms other methods in terms of SDS depletion, protein recovery, and number of protein identifications. Despite this, the need for precise pipetting challenges the reproducible success of precipitation and has prevented its wide spread use. Transmembrane electrophoresis (TME) is thus presented as a facile approach to protein purification. TME uses an electric potential to drive SDS through a dialysis membrane, while trapping protein in the sample chamber. It is amenable to both bottom-up and top-down approaches and is compatible with membrane proteins. Constant current enabled SDS depletion in 1 hour, though temperature control at constant power provides a faster rate of SDS depletion (10 min) without compromising recovery, making the device valuable to proteomic workflows.en_US
dc.identifier.urihttp://hdl.handle.net/10222/72082
dc.language.isoenen_US
dc.subjectProteomicsen_US
dc.subjectSDSen_US
dc.subjectMass Spectrometryen_US
dc.titleAssessment, Development, and Improvement of SDS Depletion Strategies in Mass Spectrometry Proteomic Workflowsen_US

Files

Original bundle

Now showing 1 - 5 of 11
No Thumbnail Available
Name:
File_S1_Acetone_High_Peptide.xls
Size:
1.35 MB
Format:
Microsoft Excel
Description:
File S1
No Thumbnail Available
Name:
File_S2_Acetone_High_Protein.xls
Size:
84.5 KB
Format:
Microsoft Excel
Description:
File S2
No Thumbnail Available
Name:
File_S3_Acetone_Low_Peptide.xls
Size:
1.43 MB
Format:
Microsoft Excel
Description:
File S3
No Thumbnail Available
Name:
File_S4_Acetone_Low_Protein.xls
Size:
127.5 KB
Format:
Microsoft Excel
Description:
File S4
No Thumbnail Available
Name:
File_S5_FASP_High_Peptide.xls
Size:
1.1 MB
Format:
Microsoft Excel
Description:
File S5

License bundle

Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.71 KB
Format:
Item-specific license agreed upon to submission
Description: