FMRP 1C3 Western Blot ( Millipore MAB2160 ) post - IP
Run tris - glycine gel .
Transfer protein to PVDF .
Rinse membrane in 1x TBST Block in 5 % non - fat milk / TBST for 30 minutes at RT .
Incubate membrane with anti - FMRP 1C3 at 1 : 4000 at RT for 1hr or O / N at 4C in 5 % non - fat milk / 1x TBST .
Rinse membrane 5x with 1x TBST over 30 minutes .
Block with 5 % IgG - Free , Protease - Free Bovine Serum Albumin in 1x TBST for 30 - 45 minutes at RT .
Add HRP - conjugated goat - anti - mouse IgG , light - chain specific antibody at 1 : 10,000 .
Incubate for 1hr at RT .
Rinse 5x with 1x TBST over 30 minutes .
Visualize with ECL solution according to manufacturer's protocol
