Deglycosylation of GTX65638-pro rat progranulin (DDDDK tagged) protein. To examine the deglycosylation of rat Progranulin (DDDDDK tagged), 1 μg of rat progranulin is denatured with 1X glycoprotein denaturing buffer at 100ºC for 10 minutes. After the addition of NP-40 and G7 reaction buffer, twofold dilutions of PNGase F are added and the reaction mix is incubated for 1 or 3 hours at 37ºC. Separation of reaction products is visualized by immunoblotting using anti-DDDDDK-HRP antibody.