Transwell Migration and Invasion Assay

Transwell migration and invasion assays were performed as described in Okimoto et al.^{35 (link)} Briefly, 8-μm-pore Matrigel coated (invasion) or non-coated (migration) Transwell inserts (BD Biosciences) were added at the top of a Transwell chamber filled with 10% FBS, RPMI media. To each insert, 2.4 × 10⁴ cells in serum-free media were added. The Transwell chambers were incubated for 20 hrs at 37 °C in the incubator. Cells that did not migrate through the pore or invade the matrigel were scraped off; the membranes were fixed in methanol for 15 min and then stained with crystal violet for 30 min. The surface of the membrane was imaged in 5 distinct fields, with a Zeiss Axioplan II immunofluorescent microscope at 10×. Invasion and migration were assessed counting the average imaged cells in the 5 regions. Results presented are from three independent experiments.

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.

Blakely C.M., Watkins T.B., Wu W., Gini B., Chabon J.J., McCoach C.E., McGranahan N., Wilson G.A., Birkbak N.J., Olivas V.R., Rotow J., Maynard A., Wang V., Gubens M.A., Banks K.C., Lanman R.B., Caulin A.F., John J.S., Cordero A.R., Giannikopoulos P., Simmons A.D., Mack P.C., Gandara D.R., Husain H., Doebele R.C., Riess J.W., Diehn M., Swanton C, & Bivona T.G. (2017). Evolution and clinical impact of co-occurring genetic alterations in advanced-stage EGFR-mutant lung cancers. Nature genetics, 49(12), 1693-1704.

Publication 2017

Transwell inserts Transwell chamber Axioplan II immunofluorescent microscope

Assays Crystal violet Immunofluorescent microscope Matrigel Membrane Methanol Serum free media

Corresponding Organization :

Other organizations : University of California, San Francisco, University College London, Cancer Research UK, CRUK Lung Cancer Centre of Excellence, The Francis Crick Institute, Stanford University, University of Colorado Denver, UCSF Helen Diller Family Comprehensive Cancer Center, Guardant (United States), Clovis Oncology (United States), University of California, Davis, University of California, San Diego

Top 5 similar protocols

Variable analysis

independent variables

Matrigel coating on Transwell inserts (coated for invasion assay, non-coated for migration assay)

dependent variables

Number of cells that migrated through the pore or invaded the Matrigel

control variables

Pore size of Transwell inserts (8-μm)
Incubation time (20 hrs)
Incubation temperature (37 °C)
Cell seeding density (2.4 × 10^4 cells per insert)
Culture media (10% FBS, RPMI)

controls

Positive control: Not explicitly mentioned.
Negative control: 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!