Works but Not the Best for Migration Assay

Duke University
Pharmacology
Research Scientist

Overall

Quality of Results

Ease-of-Optimization

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Company:

Sigma Aldrich

Product Name:

Cell Migration & Invasion Assay

Catalog Number:

ECM512

Our lab is interested in studying the role of inflammatory kinases as a therapeutic target. We perform ELISA and western blot assays to determine the role and pro-inflammatory status of macrophages in response to pharmacological inhibition of these kinases. This kit was not the best for migration assays, the positive controls were weak and the assay is not sensitive at all. It ended up only picking up gross changes, and was difficult and expensive to trouble shoot.

Experimental Design and Results Summary

Application

Migration Assay

Starting Material

N/A

Protocol Overview

Bone Marrow derived macrophages were plated .5*10^6 in each chamber in FBS free media and allowed to migrate towards 10% FBS media in the lower wells under treated conditions. Cells were allowed to migrate for 12 hours prior to analysis.

Tips

Try multiple time courses and chemoattractants.

Results Summary

Finally got this to work after multiple attempts and trying how many cells to seed and length of migration.

DOI or PMID #

N/A

Additional Notes

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Summary

The Good

Easy to seed cells in

The Bad

Not user easy, bottom well doesn't align well and splashes everywhere

The Bottom Line

Not the best but pretty much the only kit out there for this type of assay.

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