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Heavy-labeled Products (MS Standards) and Services  


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ACROBiosystems offer heavy-labeled proteins and material generation service for customers who want to do quantification of human protein biomarkers through Mass Spectrometry (MS) analysis. The proteins are expressed through 293 cells in a chemically defined environment where the protein will be labeled with [U- 13C6, 15N4]-L-Arginine and [U- 13C6, 15N2]-L-Lysine. Due to the correct post-translational modifications mechanism of human cell, the proteins will be excellent identification and quantification standards.

  BPM Cell Culture
Heavy-labeled Protein Products
Heavy-labeled Protein Services
  • Greater than 99.9% incorporation efficiency
  • Authentic posttranslational modification and protein conformation
  • Higher accuracy, higher consistency than synthetic peptide
  • Compatible with all types of MS equipments
  • Custom protein generation service available
  • Guaranteed quality and short lead-time
  • Greater than 99.9% incorporation efficiency,
    guarantee
  • 4-6 weeks lead time from sequence to end product
  • Host cell has been adapted to serum free, chemically defined SILAC medium over 10 passages
  • Avoid residual amino acid interface in traditional approach using serum contained medium


 

Comparison of ACROBiosystems protein based MS Standard Vs. Other Company peptide based MS standard
  ACRO protein Other Company peptide
Reproducibility High Low
Accuracy High low
Precision High low
Identification of best SRM and MRM transitions Yes No
PT modification and processing Yes No
Coverage >10,000s 100s

Heavy Labeled Protein Heavy Labeled Protein

Recombinant Human ADAM12 MS Standard Protein, C13 and N15-labeled (rhADAM12, Heavy Labeled) Arg 29 - Asp 513 (Accession # AAH60804.1) was produced through human 293 cells (HEK293) in chemically defined medium.

Heavy Labeled Protein Heavy Labeled Protein

Recombinant Human Apo-A1 MS Standard Protein, C13 and N15-labeled (rh Apo-A1, Heavy Labeled) Asp 25 - Gln 267 (Accession # NP_000030.1) was produced through human 293 cells (HEK293) in chemically defined medium.


 

Heavy-labeled Protein Products

Stable isotope labeling by amino acids in cell culture (SILAC) is a technique based on mass spectrometry that detects differences in protein abundance among samples using non-radioactive isotopic labeling. It is a popular method for (MS)-based quantitative proteomics. SILAC labels cellular proteomes through normal metabolic processes, incorporating non-radioactive, stable isotope containing amino acids into the proteins. The method relies on the incorporation of amino acids with substituted stable isotopic nuclei (e.g. 13C, 15N). For example, the growth medium can contain arginine labeled with six carbon-13 atoms (13C), heavy amino acid, instead of the normal carbon-12 (12C), light amino acids. Cells grown in this medium incorporate the heavy amino acids after five cell doublings and SILAC amino acids have no effect on cell morphology or growth rates, they incorporate the heavy amino acid,13C6-arginine, into all of cell proteins(Figure 1).

SILAC has emerged as a very powerful method to study cell signaling, post translation modifications such as phosphorylation, protein–protein interaction and regulation of gene expression. In addition, SILAC has become an important method in secretomics, the global study of secreted proteins and secretory pathways. It can be used to distinguish between proteins secreted by cells in culture and serum contaminants. Standardized protocols of SILAC for various application have also been published.

 

Figure 1.The principle of SILAC

Cells are differentially labeled by growing them in light medium with normal arginine (Arg-0, blue color) or medium with heavy arginine (Arg-6, red color). Metabolic incorporation of the amino acids into the proteins results in a mass shift of the corresponding peptides. This mass shift can be detected by a mass spectrometer as indicated by the depicted mass spectra. When both samples are combined, the ratio of peak intensities in the mass spectrum reflects the relative protein abundance. In this example, the labeled protein has the same abundance in both samples (ratio 1).

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If you have any questions, you can send email to proteomics@acrobiosystems.com.