Validating mGlu3 Receptor Antibody Specificity

Omission of the primary antibody and substitution with non-immune rabbit serum abolished all reactivity. Peroxidase label was not detected when bridging secondary antibodies were excluded. Blocking of biotinylated probes with avidin/biotin also abolished the signal. Finally, additional electron microscopic immunocytochemistry performed in monkey dlPFC by our group showed that the antibody used in the present study exhibited a labeling pattern that was a subset of another commercially available and well-characterized antibody (Ab6438; Abcam) raised against the shared C-terminus of mGlu2 and mGlu3 (Jin et al., 2017 (link)), consistent with the labeling of mGlu3.

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Woo E., Datta D, & Arnsten A.F. (2022). Glutamate Metabotropic Receptor Type 3 (mGlu3) Localization in the Rat Prelimbic Medial Prefrontal Cortex. Frontiers in Neuroanatomy, 16, 849937.

Publication 2022

Ab6438

Antibodies Antibody Avidin Biotin Dlpfc Electron microscopic Immune serum Immunocytochemistry Monkey Peroxidase Probes Rabbit

Corresponding Organization : Yale University

Top 5 similar protocols

Variable analysis

independent variables

Omission of the primary antibody and substitution with non-immune rabbit serum
Exclusion of bridging secondary antibodies
Blocking of biotinylated probes with avidin/biotin

dependent variables

Reactivity
Peroxidase label detection

control variables

Antibody used in the present study compared to a commercially available and well-characterized antibody (Ab6438; Abcam) raised against the shared C-terminus of mGlu2 and mGlu3

positive controls

Use of the primary antibody

negative controls

Substitution of the primary antibody with non-immune rabbit serum
Exclusion of bridging secondary antibodies
Blocking of biotinylated probes with avidin/biotin

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