Biological activities of nerve growth factor bound to nitrocellulose paper by Western blotting.
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.
Pettmann
B
B
Department of Biology, University of California San Diego, La Jolla 92093.
Manthorpe
M
M
Powell
J A
JA
Varon
S
S
D000818:Animals; D015153:Blotting, Western; D002470:Cell Survival; D002478:Cells, Cultured; D002645:Chickens; D020934:Ciliary Neurotrophic Factor; D003101:Collodion; D004591:Electrophoresis, Polyacrylamide Gel; D005726:Ganglia, Parasympathetic / Q000166:cytology; D005727:Ganglia, Spinal / Q000166:cytology; D007381:Intermediate Filament Proteins / Q000032:analysis; D008970:Molecular Weight; D009414:Nerve Growth Factors / Q000302:isolation & purification / Q000502:physiology*; D009419:Nerve Tissue Proteins / Q000502:physiology; D009474:Neurons / Q000166:cytology
D020934:Ciliary Neurotrophic Factor; D007381:Intermediate Filament Proteins; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D003101:Collodion