Transendothelial Migration Assay

The assay was described previously^{1 (link), 36 (link)}. Briefly, a monolayer of HUVECs was grown on polycarbonate filters (8 μm pore size; NeuroProbe, Gaithersburg, MD). The center 12 wells of bottom plate were filled with collagen IV. Then, melanoma cells were drawn across HUVEC monolayer with a peristaltic pump at a shear flow (0.625 or 2 dyn/cm²) for 4 hr. Migrated cells were counted using an inverted microscope (IX71, Olympus) with the NIH ImageJ software.

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.

Zhang P., Feng S., Liu G., Wang H., Fu A., Zhu H., Ren Q., Wang B., Xu X., Bai H, & Dong C. (2016). CD82 suppresses CD44 alternative splicing-dependent melanoma metastasis by mediating U2AF2 ubiquitination and degradation. Oncogene, 35(38), 5056-5069.

Publication 2016

polycarbonate filters

Assay Cells Collagen iv Melanoma Microscope Peristaltic Polycarbonate

Corresponding Organization : Southwest University

Other organizations : Shanghai Pulmonary Hospital, Tongji University, Chongqing University, Pennsylvania State University

Top 5 similar protocols

Variable analysis

independent variables

Shear flow rate (0.625 or 2 dyn/cm^2)

dependent variables

Number of migrated melanoma cells

control variables

Monolayer of HUVECs grown on polycarbonate filters (8 μm pore size)
Collagen IV filled in the center 12 wells of bottom plate
Melanoma cells drawn across HUVEC monolayer for 4 hr using a peristaltic pump
Migrated cells counted using an inverted microscope (IX71, Olympus) with NIH ImageJ software

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!