Overlay histogram showing SH-SY5Y cells stained with (green line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min. The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (, 1:25 dilution) for 60 min at 37ºC. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse lgG (166821) at 1/200 dilution for 40 min at 37ºC. Isotype control antibody (blue line) was mouse IgG1 (1μg/1x10^6 cells) used under the same conditions. Acquisition of >10, 000 events was performed.

Fluorescent image of SH-SY5Y cells stained with CHRM2 Antibody . BD-PB4096 was diluted at 1:25 dilution. An Alexa Fluor® 488-conjugated goat anti-mouse lgG at 1:400 dilution was used as the secondary antibody (green). DAPI was used to stain the cell nuclear (blue).

Immunohistochemical analysis of paraffin-embedded H. brain section using CHRM2. BD-PB4096 was diluted at 1:25 dilution. A undiluted biotinylated goat polyvalent antibody was used as the secondary, followed by DAB staining.

Immunohistochemical analysis of paraffin-embedded H. heart section using CHRM2 . BD-PB4096 was diluted at 1:25 dilution. A undiluted biotinylated goat polyvalent antibody was used as the secondary, followed by DAB staining.

Western blot analysis of lysates from SH-SY5Y cell line, human brain, mouse brain tissue(from left to right), using CHRM2 Antibodyat 1:3000 dilution was used as the secondary antibody. Lysates at 20μg per lane.