Western Blot of Mouse IgG Antibody Peroxidase Conjugated detecting mouse IgG. Mouse IgG (1.0 mg) was separated on a 4-20% gradient gel and then transferred to Whatman Protran BA 85 nitrocellulose (0.45 microm pore size) and blocked with 5% BLOTTO (p/n B501-0500) o/n at 4°C. The membrane was then probed with a 1/2,500 dilution of HRP Rabbit anti-Mouse IgG [H&L] R1253HRP for 45 min at RT. FemtoMax Super Sensitive Chemiluminescent HRP Substrate Was added and the image was captured using a BioSpectrum Imaging System with a BioChemi 500 -28C cooled 4.0 Megapixel CCD Camera from UVP (www.uvp.com, Upland, CA). Binning: 5X5. Exposure time: 2 min. Aperture open. No emission or excitation filters used.