Biological activities of nerve growth factor bound to nitrocellulose paper by Western blotting

J Neurosci. 1988 Oct;8(10):3624-32. doi: 10.1523/JNEUROSCI.08-10-03624.1988.

Abstract

We have previously developed a cell-blot technique to visualize directly in tissue extracts molecules that display the biological activity of ciliary neuronotrophic factors (CNTFs). This technique involves SDS-PAGE of the tissue extract, Western blotting on nitrocellulose paper, neuronal cell culture on the paper, and, using a vital dye, visualization of the neurons that selectively survive on the trophic factor band. In this report, we show that (1) NGF, either purified or in a crude extract from submaxillary glands, can also be successfully recognized using a slightly modified cell-blot technique; (2) a variety of ganglionic neurons can respond to distinct nitrocellulose-anchored trophic factors; (3) while CNTF and NGF can both support the survival of their common target cells, only NGF also promotes neuritic extension; and (4) both the dimeric and the monomeric forms of immobilized beta-NGF are active.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Blotting, Western
  • Cell Survival
  • Cells, Cultured
  • Chickens
  • Ciliary Neurotrophic Factor
  • Collodion
  • Electrophoresis, Polyacrylamide Gel
  • Ganglia, Parasympathetic / cytology
  • Ganglia, Spinal / cytology
  • Intermediate Filament Proteins / analysis
  • Molecular Weight
  • Nerve Growth Factors / isolation & purification
  • Nerve Growth Factors / physiology*
  • Nerve Tissue Proteins / physiology
  • Neurons / cytology

Substances

  • Ciliary Neurotrophic Factor
  • Intermediate Filament Proteins
  • Nerve Growth Factors
  • Nerve Tissue Proteins
  • Collodion