Immunofluorescence Staining of p65

Firstly, cells that grew on glass coverslips were fixed and then permeabilized with 0.25% Triton X-100 for 10 min, blocked with 0.8% BSA for 1 h at room temperature. Next, cells were incubated with the primary antibody against p65 (Cell Signaling Technology) overnight at 4 °C and treated with appropriate secondary antibody. The nucleus was counterstained with DAPI. The experiment was performed as described previously [20 (link), 21 (link)].

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Wang Y., Liu Y., Zhang M., Lv L., Zhang X., Zhang P, & Zhou Y. (2019). Inhibition of PTGS1 promotes osteogenic differentiation of adipose-derived stem cells by suppressing NF-kB signaling. Stem Cell Research & Therapy, 10, 57.

Publication 2019

primary antibody against p65

Antibody Cells Dapi Nucleus Triton x 100

Corresponding Organization :

Other organizations : Peking University

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Variable analysis

independent variables

Permeabilization with 0.25% Triton X-100 for 10 min
Blocking with 0.8% BSA for 1 h at room temperature
Incubation with primary antibody against p65 overnight at 4 °C
Treatment with appropriate secondary antibody

dependent variables

Localization and expression of p65 protein

control variables

Cells grown on glass coverslips
Fixed cells
Counterstaining of the nucleus with DAPI

controls

Positive control: Not specified
Negative control: Not specified

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