Collagenase Inhibition Assay Protocol

To measure the collagenase inhibitory activity, a weight of 1 mg of azo dye-impregnated collagen was mixed with 800 μL of 0.1 M Tris-HCl (pH 7.0), 100 μL of 200 units/mL collagenase (stock solution), and 100 μL sample and incubated at 43 °C for 1 h under shaking condition. Subsequently, the reaction mixture was centrifuged at 3000 rpm for 10 min and the absorbance of the supernatant was detected at 550 nm in a microplate reader (BioTek Synergy HT, Woburn, MA, USA).

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Wang L., Lee W., Oh J.Y., Cui Y.R., Ryu B, & Jeon Y.J. (2018). Protective Effect of Sulfated Polysaccharides from Celluclast-Assisted Extract of Hizikia fusiforme Against Ultraviolet B-Induced Skin Damage by Regulating NF-κB, AP-1, and MAPKs Signaling Pathways In Vitro in Human Dermal Fibroblasts. Marine Drugs, 16(7), 239.

Publication 2018

Synergy HT

Azo dye Collagen 1 Collagenase H condition Inhibitory Tris

Corresponding Organization : Jeju National University

Other organizations : National Institute of Biological Resources

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Variable analysis

independent variables

Sample

dependent variables

Collagenase inhibitory activity

control variables

Weight of azo dye-impregnated collagen (1 mg)
Volume of 0.1 M Tris-HCl (pH 7.0) (800 μL)
Volume of 200 units/mL collagenase (100 μL)
Incubation temperature (43 °C)
Incubation time (1 h)
Shaking condition
Centrifugation speed (3000 rpm)
Centrifugation time (10 min)
Absorbance detection wavelength (550 nm)
Microplate reader (BioTek Synergy HT, Woburn, MA, USA)

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