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Analysis of IP- MassSpec data using ProteinGURU’s Online MassSpec data Analysis Tool (OLMAT)

Analysis of IP- MassSpec data using ProteinGURU’s Online MassSpec data Analysis Tool (OLMAT). ProteinGURU.com. Step 1: Do the IP and run it on the gel.

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Analysis of IP- MassSpec data using ProteinGURU’s Online MassSpec data Analysis Tool (OLMAT)

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  1. Analysis of IP-MassSpec data using ProteinGURU’s Online MassSpec data Analysis Tool (OLMAT) ProteinGURU.com

  2. Step 1:Do the IP and run it on the gel • In this experiment, U2OS cells stably expressing GFP alone or GFP-protein fusion were treated with either DMSO or an inhibitor (drug) of interest. • The proteins were immunoprecipitated from the lysates using anti-GFP antibody and samples were run on a 4-12% gradient gel.

  3. Step 2:Cut your gel into slices and submit for Orbitrap analysis • It is important to cut the slices in the negative control lanes (GFP alone) corresponding to the MW of the slices cut from the test samples.

  4. Step 3:Archive the Orbitrap analysis files • Select all the HTML files that came from the Orbitrap analysis and compress them into a ZIP file (don’t select the folders or any other files). • For Windows: • Select all the files • Right-click • Send to • Compressed (zipped) folder • For Mac: • Select all the files • Ctrl-click • Compress (an Archive.zip file will be created)

  5. Step 4:Upload the ZIP file to OLMAT and Process the Experiment • Go to: http://www.proteinguru.com/MassSpec/OLMAT/ 2. Follow the OLMAT Demo page (3 videos). 3. Download your analysis package.

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