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Amplite universal fluorimetric mmp activity assay kit

Manufactured by AAT Bioquest
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Sourced in United States

The Amplite Universal Fluorimetric MMP Activity Assay Kit is a laboratory equipment product designed to measure the activity of matrix metalloproteinases (MMPs) in a variety of sample types. It uses a proprietary fluorescent substrate to detect MMP activity, providing a sensitive and quantitative method for assessing MMP function.

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Lab products found in correlation

Bmg fluostar optima microplate reader, by BMG Labtech (1 mentions)

3 protocols using amplite universal fluorimetric mmp activity assay kit

1

Quantifying MMP Activity in 3D Cultures

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The MMP assay was performed using the Amplite™ Universal Fluorimetric MMP Activity Assay Kit (AAT Bioquest), following the manufacturer's recommendations. 25 μL of the supernatants from the spheroid were collected after 7 days of growth and incubated with 25 μL MMP Green™ substrate working solution for 40 min. The fluorescence signal was then measured with the PHERAstar® FS multi-plate reader (λex 490 nm/λem 525 nm). Values of active MMPs were normalized to the number of surviving MB cells within the spheroid, using the ratio MMP Green/DsRed.
The same procedure was performed with the supernatants of organotypic co-cultures at experiment completion (day 14). Values of active MMPs were normalized to the number of surviving MB cells within the organotypic cultures, using the ratio MMP Green/DsRed.
Piris P., Buric D., Yamasaki T., Huchedé P., Rossi M., Matteudi M., Montero M.P., Rodallec A., Appay R., Roux C., Combes S., Pasquier E., Castets M., André N., Brémond P, & Carré M. (2023). Conditional generation of free radicals by selective activation of alkoxyamines: towards more effective and less toxic targeting of brain tumors. Chemical Science, 14(29), 7988-7998.
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2

Fluorometric Quantification of MMP Activity

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The activity of MMPs (MMP1/2/3/7/8/9/10/12/13) in culture supernatants was measured using the Amplite Universal Fluorimetric MMP Activity Assay Kit (AAT Bioquest, USA). Supernatants were activated with 4-aminophenylmercuric acetate (2 mM) overnight. Activated supernatants and controls were pipetted in duplicates into a flat-bottom culture plate (100 µl/well; Corning 96 Well White Polystyrene Microplate; Merck KGaA) and MMP Green substrate (50 µl/well; AAT Bioquest) was added. Fluorescence intensity was monitored as kinetic measurement with a plate reader (BMG FLUOstar OPTIMA Microplate Reader; BMG Labtech Inc. USA) at Ex/Em = 490/525 nm.
Seiser S., Janker L., Zila N., Mildner M., Rakita A., Matiasek J., Bileck A., Gerner C., Paulitschke V, & Elbe-Bürger A. (2021). Octenidine-based hydrogel shows anti-inflammatory and protease-inhibitory capacities in wounded human skin. Scientific Reports, 11, 32.
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3

Fluorimetric MMP Activity Assay

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MMP activity was analyzed using the Amplite Universal Fluorimetric MMP Activity Assay Kit (AAT Bioquest; Sunnyvale, CA, USA) according to the manufacturer’s instructions.
Sugano H., Shirai Y., Horiuchi T., Saito N., Shimada Y., Eto K., Uwagawa T., Ohashi T, & Yanaga K. (2018). Nafamostat Mesilate Enhances the Radiosensitivity and Reduces the Radiation-Induced Invasive Ability of Colorectal Cancer Cells. Cancers, 10(10), 386.
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