Immunofluorescent Staining of EphA2 and Shp2

Immunofluorescent staining was performed as described previously by us^{43 (link)}. Briefly, cells were plated into chamber slides (Millipore) and fixed in 4% paraformaldehyde, permeabilized, and incubated with mouse anti-EphA2 antibody (1 : 300 dilution) and rabbit anti-Shp2 antibody (1 : 100 dilution), followed by incubation with DyLight® 488 anti-mouse IgG (DI-2788, Vector Laboratories) and DyLight® 594 anti-Rabbit IgG (DI-1794, Vector Laboratories). Images were captured using an inverted confocal fluorescent microscope (LEICA TCS SP8). Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI).

Free full text: Click here

Xiang Y.P., Xiao T., Li Q.G., Lu S.S., Zhu W., Liu Y.Y., Qiu J.Y., Song Z.H., Huang W., Yi H., Tang Y.Y, & Xiao Z.Q. (2020). Y772 phosphorylation of EphA2 is responsible for EphA2-dependent NPC nasopharyngeal carcinoma growth by Shp2/Erk-1/2 signaling pathway. Cell Death & Disease, 11(8), 709.

Publication 2020

chamber slides DyLight® 488 anti-mouse IgG DyLight® 594 anti-Rabbit IgG TCS SP8

Anti antibody Anti igg Cells Confocal microscope Dapi Dilution Immunofluorescent Mouse Nuclei Paraformaldehyde Rabbit Shp2 Vector

Corresponding Organization :

Other organizations : Xiangya Hospital Central South University, Central South University, Chinese Academy of Medical Sciences & Peking Union Medical College

Top 5 similar protocols

Variable analysis

independent variables

Mouse anti-EphA2 antibody (1 : 300 dilution)
Rabbit anti-Shp2 antibody (1 : 100 dilution)

dependent variables

Localization and expression of EphA2 and Shp2 proteins

control variables

Cell culture conditions (e.g., cell line, culture medium, incubation time, temperature)
Fixation and permeabilization methods
Antibody concentrations and incubation conditions
Fluorescent dye concentrations and incubation conditions
Microscope settings and image acquisition parameters

positive controls

Not explicitly mentioned

negative controls

Not explicitly 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!