hFlt1-HEK293 lysates were immunoprecipitated by incubation with anti-HA (1 μg of antibody per 1.5 mg of protein, rabbit,, Abcam, ab9110) coupled to Protein G Dynabeads (Invitrogen, catalog number 10003D) overnight at 4°C. Proteins were eluted in sample buffer by heating to 70°C for 10 min. For Western blot analysis, equal amounts of total protein were loaded and resolved on a NuPAGE 4/12% bis/tris gel (Invitrogen) and transferred to a PVDF membrane. Blots were probed with specific antibodies for HA-tag (mouse, 1:1000, Sigma, catalog number 11583816001), Flt1 (rabbit, 1:1000, Abcam, ab3252), and anti-PFlt1 (rabbit, 1:1.000, R&D, Y1213). Enhanced chemiluminesence anti-mouse or anti-rabbit IgG conjugated with HRP (1:3000 or 1:10,000; Healthcare) was used as the secondary antibody.

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.