Osteocalcin Immunostaining of Bone Sections

Immunohistochemical staining was conducted, as previously described [41 (link), 42 (link)]. Bone segments were treated for antigen recovery through assimilation with 0.05% trypsin at 37°C for 15 minutes, and were then probed using a primary antibody against osteocalcin (Takara) overnight at 4°C. Consequently, an HRP-streptavidin recognition system (Dako) was utilized to distinguish immunoactivity, followed by counterstaining with hematoxylin (Sigma). The segments probed with polyclonal rabbit IgG (R&D Systems Inc.) acted as negative controls.

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Li L., Wang B., Li Y., Li L., Dai Y., Lv G., Wu P, & Li P. (2020). Celastrol regulates bone marrow mesenchymal stem cell fate and bone-fat balance in osteoporosis and skeletal aging by inducing PGC-1α signaling. Aging (Albany NY), 12(17), 16887-16898.

Publication 2020

hematoxylin polyclonal rabbit IgG

Antibody Antigen Bone Hematoxylin Osteocalcin Rabbit Streptavidin Trypsin

Corresponding Organization :

Other organizations : Second Xiangya Hospital of Central South University, Central South University

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Variable analysis

independent variables

Antigen recovery treatment: 0.05% trypsin at 37°C for 15 minutes

dependent variables

Immunoactivity (detected by HRP-streptavidin recognition system)

control variables

Bone segments

positive controls

Bone segments probed with primary antibody against osteocalcin (Takara)

negative controls

Bone segments probed with polyclonal rabbit IgG (R&D Systems Inc.)

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