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Bioanalysis. 2014 Nov;6(22):2969-83. doi: 10.4155/bio.14.121.

Quantification of XRCC and DNA-PK proteins in cancer cell lines and human tumors by LC-MS/MS.

Author information

  • 1Celgene Corporation, Summit, NJ 07901, USA. xwang@celgene.com.

Abstract

BACKGROUND:

The x-ray repair cross-complementing (XRCC) proteins and a catalytic subunit of nuclear DNA-dependent serine/threonine protein kinase (DNA-PK) play important roles in cancer biology. Understanding the protein expression levels allows us to reconstruct in vivo functionality and to qualify protein biomarkers.

METHODS & RESULTS:

XRCC and DNA-PK proteins in human cancer cells and tumor tissues have been identified and quantified by selected peptides using NanoLC and high-resolution mass spectrometry. The stable isotope-labeled full-length protein XRCC4 ([(13)C6, (15)N4]-arginine and [(13)C6, (15)N2]-lysine) uses as the internal standard.

CONCLUSION:

The assay range is 0.140-450 fmol (coefficient of variation: 25%) for XRCC4 in bovine serum albumen. The quantitative protein expression levels for XRCC and DNA-PK in HeLa, Ramos and HEK-293 cells and tumor tissues (lung and lymphoma) are reported.

PMID:
24785829
[PubMed - in process]
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