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. 1982;1(5):549–553. doi: 10.1002/j.1460-2075.1982.tb01207.x

Purification of a new neurotrophic factor from mammalian brain.

Y A Barde, D Edgar, H Thoenen
PMCID: PMC553086  PMID: 7188352

Abstract

We report the purification from pig brain of a factor supporting the survival of, and fibre outgrowth from, cultured embryonic chick sensory neurons. The purified factor migrates as one single band, mol. wt. 12 300, on gel electrophoresis in the presence of sodium dodecylsulphate (SDS) and is a basic molecule (pI greater than or equal to 10.1). Approximately 1 microgram factor was isolated from 1.5 kg brain. The final degree of purification was estimated to be 1.4 X 10(6)-fold, and the specific activity 0.4 ng/ml/unit, which is similar to that of nerve growth factor (NGF) using the same assay system. This factor is the first neurotrophic factor to be purified since NGF, from which it is clearly distinguished because it has different antigenic and functional properties.

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Selected References

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  1. Barde Y. A., Edgar D., Thoenen H. Sensory neurons in culture: changing requirements for survival factors during embryonic development. Proc Natl Acad Sci U S A. 1980 Feb;77(2):1199–1203. doi: 10.1073/pnas.77.2.1199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barde Y. A., Lindsay R. M., Monard D., Thoenen H. New factor released by cultured glioma cells supporting survival and growth of sensory neurones. Nature. 1978 Aug 24;274(5673):818–818. doi: 10.1038/274818a0. [DOI] [PubMed] [Google Scholar]
  3. Bocchini V., Angeletti P. U. The nerve growth factor: purification as a 30,000-molecular-weight protein. Proc Natl Acad Sci U S A. 1969 Oct;64(2):787–794. doi: 10.1073/pnas.64.2.787. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brockes J. P., Lemke G. E., Balzer D. R., Jr Purification and preliminary characterization of a glial growth factor from the bovine pituitary. J Biol Chem. 1980 Sep 25;255(18):8374–8377. [PubMed] [Google Scholar]
  5. Chapman C. A., Banks B. E., Carstairs J. R., Pearce F. L., Vernon C. A. The preparation of nerve growth factor from the prostate of the guinea-pig and isolation of immunogenically pure material from the mouse submandibular gland. FEBS Lett. 1979 Sep 15;105(2):341–344. doi: 10.1016/0014-5793(79)80644-4. [DOI] [PubMed] [Google Scholar]
  6. Edgar D., Barde Y. A., Thoenen H. Subpopulations of cultured chick sympathetic neurones differ in their requirements for survival factors. Nature. 1981 Jan 22;289(5795):294–295. doi: 10.1038/289294a0. [DOI] [PubMed] [Google Scholar]
  7. Gospodarowicz D. Purification of a fibroblast growth factor from bovine pituitary. J Biol Chem. 1975 Apr 10;250(7):2515–2520. [PubMed] [Google Scholar]
  8. Harper G. P., Thoenen H. Target cells, biological effects, and mechanism of action of nerve growth factor and its antibodies. Annu Rev Pharmacol Toxicol. 1981;21:205–229. doi: 10.1146/annurev.pa.21.040181.001225. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Oakley B. R., Kirsch D. R., Morris N. R. A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal Biochem. 1980 Jul 1;105(2):361–363. doi: 10.1016/0003-2697(80)90470-4. [DOI] [PubMed] [Google Scholar]
  12. REISFELD R. A., LEWIS U. J., WILLIAMS D. E. Disk electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature. 1962 Jul 21;195:281–283. doi: 10.1038/195281a0. [DOI] [PubMed] [Google Scholar]
  13. Server A. C., Shooter E. M. Nerve growth factor. Adv Protein Chem. 1977;31:339–409. doi: 10.1016/s0065-3233(08)60221-1. [DOI] [PubMed] [Google Scholar]
  14. Stoeckel K., Gagnon C., Guroff G., Thoenen H. Purification of nerve growth factor antibodies by affinity chromatography. J Neurochem. 1976 Jun;26(6):1207–1211. doi: 10.1111/j.1471-4159.1976.tb07008.x. [DOI] [PubMed] [Google Scholar]
  15. Suda K., Barde Y. A., Thoenen H. Nerve growth factor in mouse and rat serum: correlation between bioassay and radioimmunoassay determinations. Proc Natl Acad Sci U S A. 1978 Aug;75(8):4042–4046. doi: 10.1073/pnas.75.8.4042. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Thoenen H., Barde Y. A. Physiology of nerve growth factor. Physiol Rev. 1980 Oct;60(4):1284–1335. doi: 10.1152/physrev.1980.60.4.1284. [DOI] [PubMed] [Google Scholar]
  17. Weber K., Kuter D. J. Reversible denaturation of enzymes by sodium dodecyl sulfate. J Biol Chem. 1971 Jul 25;246(14):4504–4509. [PubMed] [Google Scholar]

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