Immunofluorescence and Western Blot Analysis of AKT Signaling

IFA and Western blotting were performed as previously described (80 (link)). For Western blotting, rabbit monoclonal anti-AKT antibody (Cell Signaling Technology, Danvers, MA), rabbit monoclonal anti-pAKT T308 antibody (Cell Signaling Technology), rabbit monoclonal anti-pAKT S473 antibody (Cell Signaling Technology), and mouse monoclonal anti-β-tubulin antibody (Sigma, St. Louis, MO) were used as primary antibodies. A rat monoclonal antibody to LANA (Abcam, Cambridge, MA) and a mouse monoclonal antibody to ORF65 (81 (link)) were used for IFA. Images were obtained using a Nikon Eclipse E400 fluorescence microscope equipped with a digital camera or a Nikon Eclipse Ti confocal laser scanning microscope (Nikon Instruments Inc., Melville, NY). Nikon NIS Elements F or Ar microscope imaging software was used for image analysis.

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Lee M.S., Yuan H., Jeon H., Zhu Y., Yoo S., Shi S., Krueger B., Renne R., Lu C., Jung J.U, & Gao S.J. (2016). Human Mesenchymal Stem Cells of Diverse Origins Support Persistent Infection with Kaposi’s Sarcoma-Associated Herpesvirus and Manifest Distinct Angiogenic, Invasive, and Transforming Phenotypes. mBio, 7(1), e02109-15.

Publication 2016

rabbit monoclonal anti-AKT antibody rabbit monoclonal anti-pAKT S473 antibody mouse monoclonal anti-β-tubulin antibody Nikon Eclipse Ti confocal laser scanning microscope

Anti antibody Antibodies Camera Confocal laser scanning microscope Fluorescence microscope Lana Microscope Monoclonal antibody Mouse Rabbit Tubulin

Corresponding Organization :

Other organizations : Eulji University, University of Southern California, University of Pennsylvania, University of Florida, Nanjing Medical University

Top 5 similar protocols

Variable analysis

independent variables

None explicitly mentioned

dependent variables

AKT protein expression and phosphorylation at T308 and S473 sites
Localization of LANA and ORF65 proteins

control variables

β-tubulin protein as a loading control for Western blotting

controls

Positive control: None mentioned
Negative control: None mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

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?

Revolutionizing how scientists
search and build protocols!