IHC Staining for CCN6 and MMP-9 Proteins

CCN6 or MMP-9 protein expression was determined by IHC staining of tissue slides^{37 (link)}. Tissue sections were deparaffinized with xylene and rehydrated with ethanol. Endogenous peroxidase activity was blocked with 3% hydrogen peroxide. Using the antigen retrieval buffer system for IHC staining, we incubated the slides with primary antibodies against CCN6 or MMP-9. The antibody binding signal was detected by the NovoLink Polymer Detection System (Leica Microsystems, Heidelberg, Germany) and visualized using 3-3’-diaminobenzidine. The sections were counterstained with hematoxylin.

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Tzeng H.E., Tang C.H., Wu S.H., Chen H.T., Fong Y.C., Lu Y.C., Chen W.C., Huang H.D., Lin C.Y, & Wang S.W. (2018). CCN6-mediated MMP-9 activation enhances metastatic potential of human chondrosarcoma. Cell Death & Disease, 9(10), 955.

Publication 2018

NovoLink Polymer Detection System

Antibodies Antibody Antigen Buffer Ccn6 Ethanol Hematoxylin Hydrogen 3 Mmp 9 Peroxidase Peroxide Polymer Protein Staining tissue Tissue Xylene

Corresponding Organization :

Other organizations : Taipei Medical University, China Medical University, Mackay Medical College, Mackay Memorial Hospital, National Yang Ming Chiao Tung University

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Variable analysis

independent variables

CCN6 protein expression
MMP-9 protein expression

dependent variables

IHC staining of tissue slides

control variables

Deparaffinization of tissue sections with xylene
Rehydration of tissue sections with ethanol
Blocking of endogenous peroxidase activity with 3% hydrogen peroxide
Use of antigen retrieval buffer system for IHC staining
Incubation of slides with primary antibodies against CCN6 or MMP-9
Detection of antibody binding signal using NovoLink Polymer Detection System
Visualization using 3-3'-diaminobenzidine
Counterstaining of sections with hematoxylin

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