This is a protocol preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Method Article
Yanbao Yu
The J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, MD 20850
Corresponding Author
Rembert Pieper
The J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, MD 20850
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
The stop-and-go-extraction tips (StageTips) have been widely used in shotgun proteomics to clean/desalt peptide samples prior to LC-MS/MS analysis. Here, an extremely simple and high throughput StageTip protocol is described. In this protocol, an adaptor is introduced to the StageTip, and makes it readily available for bench-top centrifugation. Each spin step (with 200 μL buffer loaded) takes around 2 min at 4,000 rpm. Compared with previous syringe-based manual pressure device, the spinnable StageTip is completely labor-free, automatable, and can be easily scaled up. Considered with the recently developed 96FASP method, the spinnable StageTips provide another component to the high throughput workflow for clinical proteomics and biomarker discoveries.
Keywords
Shotgun proteomics, Stop and go extraction tips (StageTips), Spinnable and automatable, Clinical proteomics, Desalt, 96FASP, Adaptor
Figure 1
Figure 2
Figure 3
Figure 4
The authors declare no competing financial interests.
Loading...
Version History
CURRENT STATUS: Posted
Version 1
Posted 08 Sep, 2014
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Associated Publications
Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips
by Juri Rappsilber, Matthias Mann, and Yasushi Ishihama
Nature Protocols (29 August, 2014)
Urine Sample Preparation in 96-Well Filter Plates for Quantitative Clinical Proteomics
by Yanbao Yu, Moo-Jin Suh, Patricia Sikorski, +3
Analytical Chemistry (10 July, 2014)
Method Article
Yanbao Yu
The J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, MD 20850
Corresponding Author
Rembert Pieper
The J. Craig Venter Institute, 9704 Medical Center Drive, Rockville, MD 20850
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 08 Sep, 2014
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
The stop-and-go-extraction tips (StageTips) have been widely used in shotgun proteomics to clean/desalt peptide samples prior to LC-MS/MS analysis. Here, an extremely simple and high throughput StageTip protocol is described. In this protocol, an adaptor is introduced to the StageTip, and makes it readily available for bench-top centrifugation. Each spin step (with 200 μL buffer loaded) takes around 2 min at 4,000 rpm. Compared with previous syringe-based manual pressure device, the spinnable StageTip is completely labor-free, automatable, and can be easily scaled up. Considered with the recently developed 96FASP method, the spinnable StageTips provide another component to the high throughput workflow for clinical proteomics and biomarker discoveries.
Figure 1
Figure 2
Figure 3
Figure 4
Loading...
Associated Publications
Protocol for micro-purification, enrichment, pre-fractionation and storage of peptides for proteomics using StageTips
by Juri Rappsilber, Matthias Mann, and Yasushi Ishihama
Nature Protocols (29 August, 2014)
Urine Sample Preparation in 96-Well Filter Plates for Quantitative Clinical Proteomics
by Yanbao Yu, Moo-Jin Suh, Patricia Sikorski, +3
Analytical Chemistry (10 July, 2014)