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A high sensitivity ZENK monoclonal antibody to map neuronal activity in Aves

Sci Rep. 2020; 
Nordmann GC, Malkemper EP, Landler L, Ushakova L, Nimpf S, Heinen R, Schuechner S, Ogris E, Keays DA.
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Gene Synthesis For the generation of a monoclonal antibody against clZENK, a 260 amino acid protein fragment (1–260) was selected as antigen; the codon-optimized mRNA was synthesized and cloned into the pET23a(+) expression vector including a C-terminal hexahistidine tag by an external contractor (GenScript USA Inc). Get A Quote

Abstract

The transcription factor ZENK is an immediate early gene that has been employed as a surrogate marker to map neuronal activity in the brain. It has been used in a wide variety of species, however, commercially available antibodies have limited immunoreactivity in birds. To address this issue we generated a new mouse monoclonal antibody, 7B7-A3, raised against ZENK from the rock pigeon (Columba livia). We show that 7B7-A3 labels clZENK in both immunoblots and histological stainings with high sensitivity and selectivity for its target. Using a sound stimulation paradigm we demonstrate that 7B7-A3 can detect activity-dependent ZENK expression at key stations of the central auditory pathway of the pigeon. Finally, ... More

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