The following primary antibodies were used for Western blotting and diluted in TBS-Tween containing 1% dry-milk or 3% BSA:
mouse anti-gephyrin (clone 3B11 cell culture supernatant, 1∶50);
rabbit anti–PSD-95 (1∶500, Synaptic Systems); rabbit anti–GABA
A-α1, guinea-pig anti–GABA
A-α5, guinea-pig anti–GABA
A-γ2 subunit (all 1∶500, Jean-Marc Fritschy, University of Zurich); rabbit anti–GABA
A-γ2 (1∶500, Synaptic Systems); mouse anti–GABA
A-β2/3 (1∶200, Millipore);
rabbit anti–β-tubulin (1∶150, Santa Cruz);
rabbit anti-GFP (1∶3,000, Abcam);
mouse anti–HA-tag (1∶1,000, clone 12CA5, Roche); and
mouse anti–myc-tag (1∶10 cell culture supernatant, clone 9E10).
HRP secondary antibodies (Santa Cruz) were used in 1∶5,000 dilutions in TBS-Tween containing 1% dry-milk. Streptavidin HRP conjugates were from Cell Signaling.
The following antibodies were used for immunofluorescence and PLA assay:
mouse anti-gephyrin (1∶50 cell culture supernatant, clone 3B11 [53] (
link));
rabbit anti-VGAT (1∶500, Synaptic Systems);
mouse anti–PSD-95 (1∶500, Thermo Scientific); rabbit anti–GABA
A-γ2 and α2 (1∶500, Synaptic Systems);
mouse anti–HA-tag (1∶1,000, clone 12CA5, Roche);
mouse anti–myc-tag (1∶10 cell culture supernatant, clone 9E10);
rabbit anti-giantin (1∶1,000, Abcam); and
rabbit anti-GFP (1∶1,000, Abcam). Secondary antibodies were all goat-raised Alexa Fluor 488 or 568 antibodies (Life Technologies).
Dejanovic B., Semtner M., Ebert S., Lamkemeyer T., Neuser F., Lüscher B., Meier J.C, & Schwarz G. (2014). Palmitoylation of Gephyrin Controls Receptor Clustering and Plasticity of GABAergic Synapses. PLoS Biology, 12(7), e1001908.
