Ab150066

The catalytic domain of Arabidopsis HMGR1S (CD1) produced in Escherichia coli was used as immunogen to produce a polyclonal antibody in rabbit and the resulting serum was immunosubstracted to remove IgG reacting against the bacterial proteins [48 (link)]. The immunopurified serum (Ab-CD1-i) was used as primary antibody at 1:500 for whole mount and 1:1000 for transmission EM. Anti-rabbit IgG secondary antibodies for HMGR detection were code Ab150066 (Abcam, Toronto, ON, Canada) coupled to Alexa Fluor-555 at 1:1000 for whole mount (Figure 1a,b), code Ab150068 (Abcam, Toronto, ON, Canada) coupled to Alexa Fluor-594 at 1:1000 for whole mount (Figure 1g), code 111-215-144 (Jackson Immunoresearch, Cambridge, UK) coupled to an 18-nm gold particle at 1:30 for transmission EM (Figure 1d) and code 111-205-144 (Jackson Immunoresearch, Cambridge, UK) coupled to a 12-nm gold particle at 1:30 for transmission EM (Figure 1e,f,l,m).
GFP was detected with Ab-5450 (Abcam, Toronto, ON, Canada) as the primary antibody at 1:1000 for whole mount and transmission EM. Anti-goat IgG secondary antibodies for GFP detection were code Ab150133 (Abcam, Toronto, ON, Canada) coupled to Alexa Fluor-488 at 1:1000 for whole mount, and code 705-215-147 (Jackson Immunoresearch, Cambridge, UK) coupled with an 18-nm gold particle at 1:15 for transmission EM.