Cellular Senescence Assay Protocol

The following antibodies were obtained from indicated suppliers: mouse
anti-BrdU FITC (BD Bioscience), mouse anti-p21 (Santa Cruz), mouse anti-BRAF (Santa
Cruz), mouse anti-β-actin (Abcam)^{19 (link)}, and mouse anti-p16 was a gift from Dr. Greg Enders.
Immunofluorescence staining and BrdU labeling for cultured cells were performed as
previously described using the antibodies listed above^{16 (link)18 (link)}. SA-β-Gal staining was
performed as previously described^{20 (link)}.

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Zeppernick F., Ardighieri L., Hannibal C.G., Vang R., Junge J., Kjaer S.K., Zhang R., Kurman R.J, & Shih I.M. (2014). BRAF mutation is associated with a specific cell-type with features suggestive of senescence in ovarian serous borderline (atypical proliferative) tumors. The American journal of surgical pathology, 38(12), 1603-1611.

Publication 2014

mouseanti-BrdU FITC mouse anti-p21 mouse anti-BRAF mouse anti-β-actin

Actin Antibodies Braf Brdu Cultured cells Fitc Immunofluorescence Mouse

Corresponding Organization : Johns Hopkins University

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Variable analysis

independent variables

Mouse anti-BrdU FITC
Mouse anti-p21
Mouse anti-BRAF
Mouse anti-β-actin
Mouse anti-p16

dependent variables

Immunofluorescence staining
BrdU labeling for cultured cells
SA-β-Gal staining

control variables

Immunofluorescence staining and BrdU labeling for cultured cells were performed as previously described
SA-β-Gal staining was performed as previously described

controls

Positive control: Not specified
Negative control: Not specified

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