Immunoblotting patterns of cytoskeletal dendritic protein expression in human neocortex

Walter E. Kaufmann, Christopher V. Taylor, Nancy A. Lishaa

Research output: Contribution to journalArticlepeer-review

Abstract

Qualitative and quantitative evaluations of cytoskeletal proteins are critical for understanding physiological and pathological processes affecting the nervous system. Most of such studies on human samples have only used immunohistochemical techniques. We describe a complementary immunoblotting approach, for the assessment of neuronal cytoskeletal proteins, which employs fresh frozen postmortem tissues. We found that cytosolic fractions are suitable for qualitative and quantitative evaluations of four major dendritic cytoskeletal proteins: microtubule-associated protein (MAP)-2, MAP-5, and high- and medium-molecular-weight nonphosphorylated neurofilaments. The enhanced chemiluminescence (ECL) technique revealed consistent and distinctive immunoblotting patterns for all four proteins in both monkey (no postmortem delay) and human (17-34 h postmortem interval) samples, some of which differed from those found in rodents. Quantitations of blots, by tissue protein-optical density curves that demonstrated linearity of the measurements in the 0- to 100-μg range, support the feasibility of these immunoassays for the study of neurologic disorders.

Original languageEnglish (US)
Pages (from-to)235-244
Number of pages10
JournalMolecular and Chemical Neuropathology
Volume31
Issue number3
DOIs
StatePublished - Aug 1997
Externally publishedYes

Keywords

  • Cerebral cortex
  • Enhanced chemiluminescence
  • Human
  • Immunoblotting
  • MAP-5
  • Microtubule-associated protein (MAP)-2
  • Neurofilaments
  • Sternberger monoclonals incorporated antibody 32 (SMI- 32)

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Clinical Neurology

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