Protein Expression and StandardsShare
Stable isotope labeled cellular biomass can be used in both proteomic and metabolomic investigations. In addition, quantitative, proteomic MS-based studies can benefit greatly from the use of purified, labeled intact protein as internal standards. The use of properly folded, labeled intact proteins are ideal internal standards because they will mimic, as close as possible, the physical and chemical properties of the target endogenous protein in a sample prior to, during and after digestion. In particular, they will undergo a similar degree of proteolytic cleavage as the unlabeled counterpart, thus improving the accuracy of the IDMS experimental result for both middle-down or bottom-up methodologies.
CIL is pleased to offer labeled cell growth media for E. coli, insect cells, yeast, and eukaryotic cells. Specific human proteins may be overexpressed in a variety of cell types using these media in conjunction with recombinant techniques so that one can obtain a relatively large amount of labeled purified protein for proteomic studies.
CIL is also please to offer reagents and kits for CellFree Sciences, which are used to produce uniform or selectively labeled proteins in yields ideal for MS-based proteomic applications.
Frequently Asked Questions
What is a bottom-up quantitative proteomic workflow?
Bottom-up proteomics is a commonly used MS-based workflow used to identify and quantitate proteins in a given sample by analyzing unique peptides generated from enzymatic cleavage of the proteins.
What protein-expression system is used to produce the heavy-labeled ApoA-1?
Why does ApoA-1 (15N) contain a his-tag?
Can I use the heavy-labeled ApoA-1 in a top-down proteomic workflow?
Zhang, C.; Gao, S.; Molascon. A.J.; Liu, Y.; Andrews, P.C. 2014. Quantitative proteomics reveals histone modifications in crosstalk with H3 lysine 27 methylation. Mol Cell Proteomics, 13(3): 749-59 PMID: 24382802
Hessling, B.; Buttner, K.; Hecker, M. 2013. Global relative quantification with liquid chromatography-matrix-assisted laser desorption ionization time-of-flight (LC-Maldi-TOF)--cross-validation with LTQ-Orbitrap proves reliability and reveals complementary ionization preferences. Mol Cell Proteomic, 12(10):2911-20. PMID: 23788530
Zhang, C.; Liu, Y.; Andrews, P.C. 2013. Quantification of histone modifications using 15N metabolic labeling. Methods, 61(3):236-43. PMID: 23454290
Kevin Millis, PhD
Senior Scientist, Application Development Manager
Kevin Millis, PhD, is the Senior Scientist and Market Development Manager for all NMR and Mass Spectrometry product lines. Kevin is responsible for Technical Services both internally and externally for all CIL customers as well as being responsible for the application and market development for the CIL products.More
Andrew Percy, PhD
Senior Applications Chemist - Mass Spectrometry
Dr. Andrew Percy is the Senior Applications Chemist for Mass Spectrometry. His responsibilities minimally involve overseeing product development, identifying new product market opportunities, assisting in the analysis of products for MS ‘omics applications, and providing technical support to customers.More
Proteomics Product Manager, Deuterated Reagents & Intermediates Product Manager, China Sales - Research Products
Tasha Agreste, Product Manager for the Proteomics product line, has been with CIL since 1989. During her tenure at CIL, Tasha has held various positions including customer service, sales and marketing, where she has proven to be instrumental in enhancing CIL’s customer relationships.More