Evaluating Anti-Inflammatory Effects of CPP-AIF

Macrophage inflammatory assay were carried out to evaluate whether CPP_AIF displayed anti-inflammation potential [22 (link),23 (link),24 (link)]. In this experiment, macrophage (Raw264.7) cells were seeded in 96-well plates (5 × 10⁵ cells/mL) and allowed to attach overnight. After attachment, cells were incubated with various concentrations of CPP_AIF for 1 h and followed by stimulation with 1 μg/mL of LPS (lipopolysaccharide). No LPS-added cells were considered as control groups. After incubation, the amount of TNF-α and IL-6 in the medium were analyzed by enzyme linked immunosorbent assay (# KHC3011 and #EH2IL6, Thermo Fisher, Waltham, MA, USA). Cell viability was measured by AlamarBlue cell viability assay (BUF012B, Bio-Rad, Hercules, CA, USA).

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Fu T.K., Kuo P.H., Lu Y.C., Lin H.N., Wang L.H., Lin Y.C., Kao Y.C., Lai H.M, & Chang M.D. (2020). Cell Penetrating Peptide as a High Safety Anti-Inflammation Ingredient for Cosmetic Applications. Biomolecules, 10(1), 101.

Publication 2020

KHC3011 #EH2IL6 AlamarBlue cell viability assay

Alamarblue Anti inflammation Assay Cell viability Cells Enzyme linked immunosorbent assay Inflammatory Macrophage Tnf α

Corresponding Organization : Industrial Technology Research Institute

Other organizations : Simpson Biotech Company (Taiwan)

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Variable analysis

independent variables

Concentration of CPP_AIF

dependent variables

Amount of TNF-α in the medium
Amount of IL-6 in the medium
Cell viability

control variables

Macrophage (Raw264.7) cells
96-well plates
Cell seeding density (5 × 10^5 cells/mL)
Incubation time (1 h)
LPS concentration (1 μg/mL)

controls

Positive control: Cells stimulated with 1 μg/mL of LPS
Negative control: Cells without LPS stimulation

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