Immunofluorescence analysis of γH2A.X

S2-013 and T3M4 cells were seeded at ~ 40% density on sterile glass cover slips in 12 well plates. The next day cells were treated with indicated amount of silibinin for 48 h. After treatment cells were washed with PBS to remove the media and immunofluorescence staining was performed as described earlier [51 (link)] using phosphorylated H2A.X (γH2A.X) antibody (1:500 dilution). Images were captured at 20X magnification using an inverted microscope (DMI600B from Leica) and processed by using Leica LAS AF software.

Free full text: Click here

Shukla S.K., Dasgupta A., Mehla K., Gunda V., Vernucci E., Souchek J., Goode G., King R., Mishra A., Rai I., Nagarajan S., Chaika N.V., Yu F, & Singh P.K. (2015). Silibinin-mediated metabolic reprogramming attenuates pancreatic cancer-induced cachexia and tumor growth. Oncotarget, 6(38), 41146-41161.

Publication 2015

DMI600B LAS AF software

Antibody Cells Dilution H2a x Immunofluorescence Microscope Silibinin Sterile

Corresponding Organization :

Other organizations : University of Nebraska Medical Center

Top 5 similar protocols

Variable analysis

independent variables

Silibinin treatment

dependent variables

Phosphorylation of H2A.X (γH2A.X)

control variables

Cell line (S2-013 and T3M4 cells)
Seeding density (~ 40% density)
Culture substrate (sterile glass cover slips)
Culture vessel (12 well plates)
Treatment duration (48 h)
Immunofluorescence staining protocol
Microscope settings (20X magnification, Leica DMI600B)

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!