GST or His fusion proteins were expressed in E. coli, purified, and eluted according to the manufacturers’ instructions (Amersham Pharmacia and QIAGEN). For hTERT fusion protein expression, 0.3 mM isopropyl-β-d-thiogalactopyranoside (IPTG) at 16°C for 20 hours was used to stimulate the expression. For other fusion proteins, bacteria were cultured for 20 hours at 20°C with 0.1 mM IPTG.Purified fusion proteins with His tag or GST tag were incubated with GST protein bound to GST beads or His protein bound to His beads for 4 hours at 4°C. After washing, the precipitated components were analyzed by immunoblot.

Note: The content above has been extracted from a research article, so it may not display correctly.



Q&A
Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.



We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.