Protocol detail

NF-κB Translocation Assay in HT29 Cells

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NF-κB translocation assay was previously described by Hajrezaie et al. In brief, HT29 cells were seeded at six-well plates on top of cover slips, and then was exposed to a BJEE IC₅₀ concentration for 3 h. One group of cells was stimulated with TNF-α (5 ng/mL) for 30 min. The staining of cells was done according to the manufacturer’s instructions. Cellomics NF-κB Activation Kit was used. The translocation of NF-κB was determined by florescence microscope (Olympus BX51).22 (link)

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Bagheri E., Hajiaghaalipour F., Nyamathulla S, & Salehen N. (2018). The apoptotic effects of Brucea javanica fruit extract against HT29 cells associated with p53 upregulation and inhibition of NF-κB translocation. Drug Design, Development and Therapy, 12, 657-671.

Publication 2018

NF-κB Activation Kit

Assay Ht29 cells Microscope Nf κb Tnf α Translocation

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Variable analysis

independent variables

BJEE treatment at IC50 concentration

dependent variables

NF-κB translocation

control variables

HT29 cell line
Exposure time of 3 hours
TNF-α stimulation at 5 ng/mL for 30 minutes

positive controls

TNF-α stimulation (5 ng/mL for 30 minutes)

negative controls

No treatment (untreated cells)

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