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Bioassay, isolation and studies on the mechanism of action of neurite extension factorThe identification and purification of molecules active in promoting neurite outgrowth requires a sensitive reproducible bioassay. A quantitative bioassay was utilized to purify a neurite extension factor (NEF) based on counting the number of phase bright neurons with processes at least equal to one cell body diameter after 20 hrs. in culture is defined, serum free medium. Using a combination of heat treatment DEAE cellulose chromatography and gel filtration, an acidic protein of M sub r = 75,000 was highly purified. Upon reduction, it yields subunits of M sub r = 37,000. Purified fractions are active half maximally at 100 ng/ml in inducing neurite outgrowth in this bioassay. Currently, monoclonal antibodies to NEF are being produced. Female Balb C mice were immunized with the antigen and fusions with mouse myeloma cells will be performed to yield hybridoma cells.
Document ID
19840010813
Acquisition Source
Legacy CDMS
Document Type
Conference Paper
Authors
Kligman, D.
(National Inst. of Mental Health Bethesda, MD, United States)
Date Acquired
August 12, 2013
Publication Date
February 1, 1984
Publication Information
Publication: NASA. Washington NASA Space Biol. Program
Subject Category
Life Sciences (General)
Accession Number
84N18881
Distribution Limits
Public
Copyright
Work of the US Gov. Public Use Permitted.

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