Jeffrey J. Porter, Justin Wildsmith, Christopher D. Melm, mark D. Schuchard, Kevin M. Ray, Dian Er Chen, Graham B.I. Scott
You are watching: Most abundant protein in blood plasma
AQUA™ Depletion plasma facts
Figure 2.Plasma Protein Depletion Workflow
High-Abundance Protein Depletion
An AQUA™ Peptide stock solution was prepared by dissolve isotopically labeled versions of the two target peptides (FQNALLVR* and GASQAGAPQGR*, respectively) in 0.1% TFA come a final concentration that 62.5 fmol/mL. Each digest sample (whole and depleted plasma) was dissolved in 20 mL that the AQUA™ Peptide share solution. Samples to be analyzed making use of an Agilent capillary 1100 HPLC coupled to a Thermo Finnigan LTQ straight ion catch mass spectrometer. Utilizing an LC-SRM method, the absolute quantity of every peptide (and corresponding protein) was determined.
Figure 3. Overview that the Protein-AQUA™ Method. This technique was emerged by Dr. Steve Gygi and colleagues in ~ Harvard medical School
Select one optimal tryptic peptide and also stable isotope amino mountain from the sequence of your protein the interest.
Order artificial AQUA™ Peptide from.
Optimize LC-MS/MS separation protocol because that quantitation.
Extract protein from biological samples and include known quantity of AQUA™ peptide.
Analyze by LC-MS/MS or MALDI to quantitate protein that interest.
See more: What Does Run A Train On You Mean, Alexandria Ocasio
Click on picture to enlarge.
Anderson NL, Anderson NG. 2002. The human Plasma Proteome. Mol cell Proteomics. 1(11):845-867. Http://dx.doi.org/10.1074/mcp.r200007-mcp200
Gerber SA, rush J, Stemman O, Kirschner MW, Gygi SP. 2003. Absolute quantification that proteins and also phosphoproteins from cabinet lysates through tandem MS. Proceedings of the nationwide Academy that Sciences. 100(12):6940-6945. Http://dx.doi.org/10.1073/pnas.0832254100