Recombinant Anti-Gephyrin antibody [EPR12650] (ab181382)

Lanes 1 - 4: Merged signal (red and green). Green - ab181382 observed at 90 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab181382 was shown to recognize Gephyrin in wild-type HAP1 cells as signal was lost at the expected MW in GPHN (Gephyrin) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GPHN (Gephyrin) knockout samples were subjected to SDS-PAGE. Ab181382 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue sections labeling Gephyrin with purified ab181382 at 1/100 dilution (1.62 µg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

Immunohistochemical analysis of parafiin embedded human brain tissue using ab181382 (unpurified).

Heat mediated antigen retrieval was performed with citrate buffer pH 6 before commencing with IHC staining protocol.