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. 1985 Jan 7;53(1):115-9.
doi: 10.1016/0304-3940(85)90106-5.

Purification of gamma-enolase messenger ribonucleic acid from rat brain by an immunoadsorption method

O Nybroe, O C Hansen, E Bock

Purification of gamma-enolase messenger ribonucleic acid from rat brain by an immunoadsorption method

O Nybroe et al. Neurosci Lett. .

Abstract

Messenger ribonucleic acid (mRNA) coding for the brain-specific protein gamma-enolase was isolated by an immunopurification procedure. Rat brain polysomes including nascent polypeptide chains were reacted with specific gamma-enolase antibody. The polysome-antibody complexes were subsequently adsorbed to protein A-Sepharose. After extensive washing, RNA was eluted and applied to an oligo(dT)-cellulose column. Purified mRNA was translated in vitro in a mRNA-dependent rabbit reticulocyte lysate system. The synthesized product was identical to gamma-enolase synthesized by free polysomes from rat brain. Immunoisolated gamma-enolase mRNA was enriched 380-fold compared to total mRNA extracted from free polysomes. This result indicates that low-abundance mRNAs may conveniently be isolated from brain tissue by immunoadsorption of polysomes.

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