Rat anti ha hrp

Antibodies were added to Costar 96-well half area plates (Corning, Corning, NY, USA Product No. 3690) coated with 1 μg/mL of the antigen. After incubation at room temperature for 1 hour, the plates were washed three times with TBST and incubated with rabbit anti-human antibody-horseradish peroxidase (HRP) (Pierce, Rockford, IL, USA) for IgG antibodies and rat anti-HA-HRP (Clone F10, Roche Life Science, Basel, Switzerland) for Fab antibodies, respectively. Plates were washed three times, tetramethylbenzidine (TMB) peroxidase substrate (SurModics, Eden Prairie, MN, USA) was added and reactions were stopped by adding 1 N sulfuric acid (DukSan, Ansan, Korea). Absorbance at 450 nm was measured using a Victor X3 instrument (PerkinElmer, Waltham, MA, USA).

Mouse anti-SCAR (DSHB P1C1-SCAR, RRID: AB_2618386) was used at a dilution 1:250. Rat anti HA-HRP (Roche catalog number: 12013819001, RRID:AB_390917) was used at a dilution of 1:500. Guinea Pig anti-Non-stop (Mohan et al., 2014a (link)) was used at a dilution of 1:1000. Guinea Pig anti-Ada2b (Kusch et al., 2003 (link)) was used at a dilution of 1:1000. Rabbit anti-Atxn7 (Mohan et al., 2014a (link)) was used at a concentration of 1:2000. Goat anti Guinea Pig HRP (Jackson ImmunoResearch INC catalog number: 106-035-003, RRID:AB_2337402) was used at a dilution of 1:10,000. Goat anti-mouse HRP (Jackson ImmunoResearch INC catalog number: 115-035-003, RRID:AB_10015289) was used at a dilution of 1:5000. Goat anti-Rabbit HRP (Jackson ImmunoResearch INC catalog number: 111-035-003, RRID:AB_2313567) was used at a dilution of 1:10,000.