Western Blot Analysis of Cell Signaling

Western blots were performed as described previously^{19 (link),20 (link),47 (link)} using NuPage 4–12% gels and the following antibodies. Uncropped, original blots are provided in Supplementary Fig. 1.
The following primary antibodies were used: anti-TRPV4 antibody (raised in mouse; 1B2.6; Millipore Sigma; MABS466; 1:1,000), anti-ARP3 (mouse; FMS338; Abcam; ab49671; 1:5,000), anti-ARPC4 (rabbit; Abcam; ab217065; 1:2,000), anti-integrin β1 antibody (rabbit; Cell Signaling; 4706S; 1:1,000) and anti-NHE1 (raised in mouse; 54; Santa Cruz Biotechnology; sc-136239; 1:200). GAPDH was used as a loading control (rabbit; 14C10; Cell Signaling; 2118S; 1:5,000).
The following secondary antibodies were used: anti-mouse IgG, HRP-linked antibody (Cell Signaling; 7076S; 1:1,000) and anti-rabbit IgG, HRP-linked antibody (Cell Signaling, 7074S; 1:1,000).

Free full text: Click here

Bera K., Kiepas A., Godet I., Li Y., Mehta P., Ifemembi B., Paul C.D., Sen A., Serra S.A., Stoletov K., Tao J., Shatkin G., Lee S.J., Zhang Y., Boen A., Mistriotis P., Gilkes D.M., Lewis J.D., Fan C.M., Feinberg A.P., Valverde M.A., Sun S.X, & Konstantopoulos K. (2022). Extracellular fluid viscosity enhances cell migration and cancer dissemination. Nature, 611(7935), 365-373.

Publication 2022

MABS466 sc-136239 anti-rabbit IgG, HRP-linked antibody

Anti antibody Anti igg Antibodies Antibody Gapdh Gels Integrin Mouse Nhe1 Rabbit Trpv4 Western blots

Corresponding Organization :

Other organizations : Johns Hopkins University, Binghamton University, National Cancer Institute, National Institutes of Health, Center for Cancer Research, Pompeu Fabra University, University of Alberta, Carnegie Institution for Science, Department of Embryology, Auburn University, Sidney Kimmel Comprehensive Cancer Center

Top 5 similar protocols

Variable analysis

independent variables

Western blot protocol (described previously in references 19, 20, and 47)

dependent variables

Protein expression of TRPV4, ARP3, ARPC4, integrin β1, and NHE1

control variables

GAPDH (used as a loading control)

controls

Uncropped, original blots are provided in Supplementary Fig. 1 (positive control)

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!