Protocols we use in the facility (updated May 2017)
- Tissue and Cell lysis protocol
- Protease Max In-Gel Digestion Protocol
- In Solution Digestion Protocol
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Using A SDS Page Gel to clean up an IP experiment
Easy way to clean up an IP experiment
- HyperSep Tip SPE
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Digestion Proteins off Magnetic Beads
You can use this to Digest proteins right off magnetic beads
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Using Polyacrylamide to clean up a sample
No need to even run a gel, watch out for Acrylamide modifications with this one!
- SDS Page
- Pulling and Packing your own nanospray columns
- Using iRT retention time standards in Skyline
Other Nice Protocols
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This is another good ingel digestion protocol
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Magnetic Bead IP and Digestion
works great!
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Immunoprecipitation protocols and Tips
From Abcam
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From the Laboratory of Mass Spectrometry and Gaseous Ion Chemistry
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Nice document comparing various protein precipitation methods
- Plant Protein Extraction that usually works 🙂
- Good Affinity Purification Nature Protocols paper
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Protocols from other People that we find work well
Protocols from Other sources
I have not tried all of these, but they seem good. Let me know if they work for you. The Shared proteomics links seem not to be working at the moment..Sorry.
- Duke Proteomics Core Facility
They seem to have some really nice protocols here -
From Shared Proteomics
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From Shared Proteomics
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Destaining, reduction and alkylation
From Shared Proteomics
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from Shared Proteomics
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From Shared Proteomics
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From Shared Proteomics
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Preparation of HeLa peptides for LC-MS
From Protocol Exchange
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Peptide Fractionation and Clean-up Protocols
From the UWPR Proteomics Resource Center
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Great protocol for Plant tissue
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Good Protein extraction and digestion protocol for HEK cells
This is from Chris Collangelo @ Yale and was used with great success on the ABRF sPRG 2013 study
- Everything Dynabeads
- Protocols from UWPR
Software Tutorials
