Spectrophotometric assessment of nuclear proteins: a preliminary study

Eur J Histochem. 2004 Jul-Sep;48(3):329-34.

Abstract

Qualitative evaluation of protein content in formalin fixed, paraffin-embedded tissues is usually performed by means of cytofluorimetric analysis. On the other hand, several studies underline the opportunity to measure the concentration of nuclear proteins, which is often accomplished by using complex techniques and instrumentation. In the present work, we suggest a new application for the spectrophotometric evaluation of protein content on extracted and isolated nuclei, based on EDTA treatment of specimens and chemical extraction of proteins, followed by direct spectrophotometric measurement at UV wavelengths. We also demonstrate how this parameter correlates with other diagnostic factors, such as the proliferation index (MIB-1) and the DNA content (ploidy) of cells. This method is simple and effective, yet less expensive than other protein quantitation protocols.

MeSH terms

  • Breast Neoplasms / chemistry*
  • Breast Neoplasms / pathology
  • Cell Nucleus / chemistry
  • Cell Nucleus / pathology
  • DNA, Neoplasm / analysis
  • DNA, Neoplasm / isolation & purification
  • Female
  • Fixatives / chemistry
  • Flow Cytometry
  • Formaldehyde / chemistry
  • Humans
  • Immunohistochemistry
  • Ki-67 Antigen / analysis
  • Nuclear Proteins / analysis*
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / isolation & purification
  • Paraffin Embedding
  • Ploidies
  • Spectrophotometry / methods*
  • Tissue Fixation

Substances

  • DNA, Neoplasm
  • Fixatives
  • Ki-67 Antigen
  • Nuclear Proteins
  • Formaldehyde