The use of mass spectrometry for the characterization of individual or complex protein samples continues to be one of the fastest growing fields in the life science market.
Bottom-up proteomics is the traditional approach to address these questions. Optimization of each the individual steps (e.g. sample prep, digestion and instrument performance) is critical to the overall success of the entire experiment.
To address issues that may arise in your experimental design, Promega has developed unique tools and complementary webinars to help you along the way.
Here you can find a summary of individual webinars for the following topics: Continue reading “Bottom-up Proteomics: Need Help?”
The novel mass spectrometry compatible surfactant sulfonate-(sodium 3-((1-(furan-2-yl)undecyloxy) carbonylamino)-propane-1-sulfonate (i.e.ProteaseMAX) facilitates both in-gel and in-solution digestion applications by reducing the time required, enabling protein solubilization/denaturation and increasing peptide/protein identifications.
A new application was highlighted in a recent publication (1) which utilized ProteaseMAX to lyse cells prior to trypsin digestion and subsequent mass spec analysis. The composition of the buffer determines the overall efficiency of cell lysis, dissociation of protein complexes, protein solubility and ease of removal prior to LC/MS-MS analysis.
When compared to lysis buffers containing either urea or SDC, ProteaseMAX provided the optimal number of identified peptides/proteins.
In addition it can be easily removed from the lysate by acidic precipitation.
- Pirmoradian, M. et al. (2013). Rapid and deep human proteome analysis by single-dimension shotgun proteomics. Mol. Cell. Prot. 12, 3330–8.
In-gel digestion complete in only 1 hour.[/caption]Identification of proteins resolved by SDS-PAGE requires a lengthy in-gel digestion and extraction procedure. The publication, Mass Spectrometry Compatible Surfactant for the Optimized In-Gel Protein Digestion, Saveliev, S. et al. (2013) Anal. Chem. 85, 907–14, addressed these obstacles by illustrating the technical benefits of the ProteaseMAX™ mass spectrometry surfactant. A recent webinar reviewed key aspects of that paper that included:
Improved peptide recovery from gels: ProteaseMAX™ improved identification of proteins by enhanced protein digestion, increased peptide extraction and minimized postdigestion peptide loss. This is a major benefit for the in-gel digestion of low abundant proteins and enables the use of minimal sample material.
The webinar also contained information regarding the how the structure of ProteaseMAX™ Surfactant enables it to compatible with mass spec and how it can be used for protein denaturation and solubilization.
About the Webinar Series
www.promega.com/webinars/ provides a schedule of upcoming webinars. In addition, there are links to previous webinars, which allow you to view the recording or download a pdf of the presentation. There is also a pdf of additional material available for each past webinar.
To register for a webinar, use the Registration link at: www.promega.com/webinars/ This allows you to view a live webinar and participate in the live chat.
Need a reminder? You can also sign-up for monthly invitations to webinars at the webinars page (see link above). Note: Live chat is only available for live webinars, not recorded webinars.