Samples were separated by an 8% SDS-PAGE. Proteins were transferred to a nitrocellulose membrane (Protan BA85, 0.45 μm, GE Healthcare) at 300 mA for 2.5 h. The membranes were blocked in 5% milk (skim milk powder, LP0031, Oxiod) in 1× PBS (P1379, Sigma-Aldrich), incubated with a primary antibody diluted in 5% milk in 0.1% PBS-Tween 20 (PBST) for 1 h, washed three times for 10 min in 0.1% PBST, incubated with the secondary antibody diluted in 5% milk in 0.1% PBST for 30 min, and washed three times again in 0.1% PBST. The signal was detected using direct imaging by the Odyssey Classic imager (LI-COR). Intensity of bands was quantified using the Image Studio software. Unmodified blots corresponding to key experiments presented in Figs. 1 –8 can be found in Supplementary Figs. 10 and 11 .
Protan ba85
Manufactured by GE Healthcare
The Protan BA85 is a laboratory filtration device designed for the separation of biomolecules, such as proteins and nucleic acids, from complex mixtures. It utilizes a nitrocellulose membrane to facilitate the filtration process. The core function of the Protan BA85 is to provide a reliable and efficient means of sample preparation for further analysis or purification.
Automatically generated - may contain errors
2 protocols using protan ba85
1
Western Blot Analysis Protocol
2
SDS-PAGE and Immunoblot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SDS-PAGE was performed as described previously (Laemmli, 1970 (link)) using precasted Mini-PROTEAN TGX 4–20% acrylamide gels (Bio-Rad). For immunoblot, nitrocellulose membranes (Protan BA85; GE Healthcare) were probed with anti-HA (Anti-HA-Peroxidase High Affinity 3F10; Roche), peroxidase anti-peroxidase (Sigma-Aldrich) or anti-GFP (monoclonal antibody JL-8; Living Colors), followed by a peroxidase-conjugated secondary goat anti-rabbit IgG antibody (Sigma-Aldrich). Protein transfer, blotting, and chemiluminescence detection were performed using standard procedures. Detection of proteins was performed using the ECL kit (GE Healthcare).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!