Immunohistochemistry and Co-IP Protocol for WWOX/WOX1

IHC, co-immunoprecipitation and Western blotting were carried out, as described [12 (link)–15 (link), 17 (link)]. Antibody against WWOX/WOX1 was used for co-immunoprecipitation [12 (link)–15 (link), 17 (link), 22 (link)]. For IHC, skin tissue sections from patient were immersed in blocking solution (2% BSA) for 1 hr, followed by adding an aliquot of antibody against WWOX/WOX1 or TRAF2 for 2 hr incubation at room temperature. Following washing with PBS for 3 times, the tissue sections were incubated with aliquots of horseradish peroxidase (HRP)-conjugated secondary antibodies, and the resulting color was then developed using an immunohistochemical staining kit (Dako). In controls, no primary antibodies were used. The images were analyzed by an NIH Image J program.

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Chen S.J., Lin P.W., Lin H.P., Huang S.S., Lai F.J., Sheu H.M., Hsu L.J, & Chang N.S. (2015). UV irradiation/cold shock-mediated apoptosis is switched to bubbling cell death at low temperatures. Oncotarget, 6(10), 8007-8018.

Publication 2015

immunohistochemical staining kit

Antibodies Antibody Co immunoprecipitation Horseradish peroxidase Patient Skin tissue Tissue Traf2 Wwox

Corresponding Organization :

Other organizations : National Cheng Kung University, Chi Mei Medical Center, New York State Office for People With Developmental Disabilities, SUNY Upstate Medical University

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Variable analysis

independent variables

Antibody against WWOX/WOX1 used for co-immunoprecipitation

dependent variables

Protein-protein interactions detected by co-immunoprecipitation
Protein expression levels detected by Western blotting
Protein localization and expression patterns detected by IHC

control variables

Experimental procedures for IHC, co-immunoprecipitation, and Western blotting as described in references [12]-[15], [17], and [22]
Blocking solution (2% BSA) used for IHC tissue sections
Horseradish peroxidase (HRP)-conjugated secondary antibodies used for IHC
Immunohistochemical staining kit (Dako) used for IHC

positive controls

No positive controls were explicitly mentioned.

negative controls

For IHC, tissue sections incubated without primary antibodies were used as negative controls.

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