Use of Reverse Phase Protein Microarrays to Study Protein Expression in Leukemia: Technical and Methodological Lessons Learned

Steven M. Kornblau, Kevin R. Coombes

Research output: Chapter in Book/Report/Conference proceedingChapter

24 Scopus citations

Abstract

Leukemias are well suited to proteomic profiling by RPPA due to the ready accessibility of blasts from the blood or marrow. In this review, we review methodological and procedural issues that affect the quality of RPPA data. We recommend contact printers that minimize sample quantities and evaporation and maximize sample per slide. The impact of sample selection and handling is reviewed as well. Protein is best prepared fresh on the date of acquisition as cryopreservation changes protein expression levels in some diseases. Rapid processing is also required to avoid changes in phosphorylation over time. Sample source, blood vs. marrow does not seem to affect results as long as leukemic blast enrichment procedures are utilized. The choice of the correct “normal” control is important for comparing diseased to “normal” expression. Various means of normalizing the data are discussed.

Original languageEnglish (US)
Title of host publicationProtein Microarrays
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages141-155
Number of pages15
ISBN (Print)9781617792854
DOIs
StatePublished - 2011
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume785
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Leukemia
  • Proteomics
  • RPPA
  • Reverse phase protein array

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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