Western blot of E.coli whole cell extract transfected with GST epitope tagged human ATF3. GeneTexs Affinity purified anti-ATF3 detects a band ~48 kDa corresponding to recombinant human ATF3. Immunostaining using GeneTexs 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 4oC. 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. LICORs OdysseyR Infrared Imaging System was used to scan and process the image. Other detection systems will yield similar results.