Explanted TEVG samples were fixed in 10% formalin for 24 hours at 4°C, and then embedded in paraffin. For the standard histology, staining of tissue sections with hematoxylin and eosin, Masson trichrome, Verhoeff-Van Gieson, and von Kossa stains were done. For the immunohistochemistry, we used the primary antibodies including von Willebrand factor (vWF) (1:2000; Dako, Cat:A0082), α-smooth muscle actin (SMA) (1:500; Abcam, Cat:ab5694), and CD68 (1:200, Abcam,ab31630). Detection of the antibody binding was done by using biotinylated secondary antibodies (Vector Laboratories, Burlingame, Calif), followed by incubation with streptavidinated horseradish peroxidase (Vector Laboratories). The chromogenic reaction with 3,3-diaminobenzidine (Vector Laboratories) was performed for the development of the immunohistochemistry. Counterstaining of the Nuclei was done with Gill’s hematoxylin (Vector Laboratories).
Streptavidinated horseradish peroxidase
Manufactured by Vector Laboratories
Streptavidinated horseradish peroxidase is an enzyme-conjugated reagent used in various bioassay and detection applications. It consists of the enzyme horseradish peroxidase covalently attached to streptavidin, a protein that binds strongly to the vitamin biotin. This reagent can be utilized in procedures involving biotin-based detection systems.
Automatically generated - may contain errors
2 protocols using streptavidinated horseradish peroxidase
1
Histological and Immunohistochemical Characterization of TEVG
2
Histological Analysis of Grafts
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Chronic phase grafts were explanted, fixed in 10% formalin for 24 hours, and then embedded in paraffin. Standard histology was completed using hematoxylin and eosin stains. Immunohistochemistry was performed using α-smooth muscle actin (αSMA; 1:1600 M0851 Dako). Detection of antibody binding was performed using biotinylated secondary antibodies (Vector Laboratories), followed by incubation with streptavidinated horseradish peroxidase (Vector Laboratories). A chromogenic reaction with 3,3-diaminobenzidine (Vector Laboratories) was performed for the development of immunohistochemistry. Counterstaining of the nuclei was performed with Gill's hematoxylin (Vector Laboratories).
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