Visualizing Melanoma Cell Actin Cytoskeleton

Coverslips were coated with warm Oregon Green® 488-conjugated gelatin (Invitrogen) as described in Ref(9 (link)). Melanoma cells (7.5 × 10⁴) were seeded on the coverslips and after 16-hour incubation at 37°C they were fixed in 4% paraformaldehyde in PBS. After permeabilization in 0.05% triton x-100 in PBS, cells were stained with rhodamine-conjugated phalloidin (Invitrogen, to visualize actin) and hoechst (Thermo Scientific, to visualize nuclei). Coverslips were mounted onto glass slides with aqua-mount media (Polysciences, Warrington, PA). Fluorescent images were captured using a Nikon TE2000-E inverted microscope equipped with Roper CoolSnap HQ CCD camera and MetaVue 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.

Bianchi-Smiraglia A., Bagati A., Fink E.E., Moparthy S., Wawrzyniak J.A., Marvin E.K., Battaglia S., Jowdy P., Kolesnikova M., Foley C.E., Berman A.E., Kozlova N.I., Lipchick B.C., Paul-Rosner L.M., Bshara W., Ackroy J., Shewach D.S, & Nikiforov M.A. (2016). Microphthalmia-Associated Transcription Factor Suppresses Invasion by Reducing Intracellular GTP Pools. Oncogene, 36(1), 84-96.

Publication 2016

Oregon Green® 488-conjugated gelatin rhodamine-conjugated phalloidin hoechst aqua-mount media

Actin Cells Gelatin Melanoma Microscope Nuclei Oregon green 488 Paraformaldehyde Rhodamine phalloidin Triton x 100

Corresponding Organization :

Other organizations : Roswell Park Comprehensive Cancer Center, Institute of Biomedical Chemistry, University of Michigan–Ann Arbor

Top 5 similar protocols

Variable analysis

independent variables

Coating the coverslips with warm Oregon Green® 488-conjugated gelatin

dependent variables

Melanoma cell behavior and morphology

control variables

Incubation time (16 hours)
Incubation temperature (37°C)
Fixation in 4% paraformaldehyde in PBS
Permeabilization in 0.05% triton x-100 in PBS
Staining with rhodamine-conjugated phalloidin and hoechst
Mounting coverslips onto glass slides with aqua-mount media

controls

Positive control: None explicitly mentioned
Negative control: None 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!