Application
Western blot of E.coli whole cell extract transfected with GST epitope tagged human ATF3. GeneTex's Affinity purified anti-ATF3 detects a band ~48 kDa corresponding to recombinant human ATF3. Immunostaining using GeneTex's anti-GST epitope tag antibody confirms the composition of the recombinant band (not shown). The protein was transferred to nitrocellulose using standard methods. After blocking with 5% goat serum and 0.5% non fat milk in PBS, the membrane was probed with the primary antibody diluted 1:200 in 0.2X blocking buffer in PBS overnight at 4°C. Reaction was followed by washes and reaction with a 1:5000 dilution of IRDye800 conjugated Gt-a-Rabbit IgG [H&L] for 30 min at room temperature. LICOR's Odyssey® Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results.
Application
Western blot of mammalian whole cell extract transfected with HA epitope tagged human ATF3. GeneTex's Affinity purified anti-ATF3 detects a band ~31 kDa corresponding to recombinant human ATF3. Immunostaining using GeneTex's anti-HA epitope tag antibody confirms the composition of the recombinant band (not shown). The protein was transferred to nitrocellulose in 30 minutes using standard methods. After blocking with 5% goat serum and 0.5% non-fat milk in PBS, the membrane was probed with the primary antibody diluted 1:200 in 0.2X blocking buffer in PBS overnight at 4°C. Reaction was followed by washes and reaction with a 1:5000 dilution of IRDye™800 conjugated Gt-a-Rabbit IgG [H&L] for 30 min at room temperature. LICOR's Odyssey® Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results.