Subcellular fractionation and enrichment strategies for improved mass spectrometry analysis
While proteomic researchers can identify thousands of proteins in a single sample, many of the biologically significant proteins are often not detected by the mass spectrometer due to their very low abundance amidst the sheer number of total proteins present in the sample. Fractionation or simplification of a complex sample before mass spec analysis helps bring many of these lower copy number proteins into view, helping researchers better understand changes in their expression, cellular location, post-translational modifications, and some of the interacting proteins that may influence their biological function. Fractionation can be accomplished on multiple levels, ranging from very broad subcellular compartmentalization to targeted immunoselection.
In this webinar, you will learn how:
- Differential detergent subcellular fractionation can improve protein identification and identify co-localizing proteins.
- Enrichment of integral and peripheral membrane proteins is accomplished using a non-denaturing differential detergent solubilization protocol.
- Immunoprecipitation facilitates the identification of proteins that otherwise cannot be detected in whole cell lysates, and how IP in combination with SILAC facilitates monitoring changes in protein expression, post-translational modifications, and identify potential interacting proteins