PMID: 8450192Apr 1, 1993

Brain-derived neurotrophic factor (BDNF) peptide antibodies: characterization using a Vaccinia virus expression system

The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society
C WetmoreL Olson

Abstract

We describe and characterize a series of polyclonal antibodies, generated against amino acid sequences unique to various regions within pro- and mature brain-derived neurotrophic factor (BDNF), a member of the highly conserved nerve growth factor (NGF) family of neurotrophins. Synthetic peptides were coupled to carrier proteins in the presence of glutaraldehyde to restrict the host animals' immune response to epitopes that are compatible with aldehyde fixation. Initial screenings of the reactivity of the antisera were made on brain sections processed for immunohistochemistry after peptide injections into brain parenchyma. As a means of further characterizing these peptide antisera, we have evaluated the reactivity and specificity of the peptide antibodies in BHK cells expressing recombinant pro- and mature BDNF protein from a T7 RNA polymerase-driven Vaccinia virus system. Several of the antibodies strongly stained components of cells transfected with the BDNF gene but did not label wild-type cells nor cells containing only the expression vector. It has also been possible to detect differential compartmentalization of the BDNF protein at various stages of processing in the BHK cells, as well as in situ in cryostat sections of b...Continue Reading

References

Oct 31, 2002·The European Journal of Neuroscience·L A KarchewskiV M K Verge
Sep 1, 1995·The European Journal of Neuroscience·M M Dugich-DjordjevicF Hefti

Related Concepts

Buffalopox virus
Transfection
Proteins, Recombinant DNA
Immune Sera
Immunofluorescence Microscopy
Immunocytochemistry
Brain-Derived Neurotrophic Factor
Neurite Outgrowth Factors
Peptide Fragments
Brain Chemistry

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