Immunohistochemical Analysis of Ki-67 and STAT5A

Immunohistochemistry analysis was performed on the paraffin-embedded tumor sections as previously described 29 (link). After antigen retrieval and blocking with normal serum, the slides were incubated with Ki-67 antibody (Dako, Mississauga, ON, Canada) or STAT5A antibody (Proteintech) overnight at 4 ºC, washed with phosphate buffered saline, followed by biotinylated secondary antibody and peroxidase-conjugated avidin-biotin complex (Dako). Immunostaining was visualized using 3,3'-diaminobenzidine (Dako), which served as a chromogen, and the sections were counterstained with hematoxylin. Ki-67 proliferation index and staining scores for STAT5A were determined as described previously 25 (link), 30 (link). For STAT5A, specimens assigned scores of 0~1 were categorized as low expression, whereas scores 2~3 were determined as high expression.

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Li B., Hong P., Zheng C.C., Dai W., Chen W.Y., Yang Q.S., Han L., Tsao S.W., Chan K.T., Lee N.P., Law S., Xu L.Y., Li E.M., Chan K.W., Qin Y.R., Guan X.Y., Lung M.L., He Q.Y., Xu W.W, & Cheung A.L. (2019). Identification of miR-29c and its Target FBXO31 as a Key Regulatory Mechanism in Esophageal Cancer Chemoresistance: Functional Validation and Clinical Significance. Theranostics, 9(6), 1599-1613.

Publication 2019

Ki-67 antibody peroxidase-conjugated avidin-biotin complex 3,3'-diaminobenzidine

Antibody Antigen Avidin Biotin Chromogen Embedded paraffin Hematoxylin Immunohistochemistry Peroxidase Phosphate Saline Serum Stat5a Tumor

Corresponding Organization :

Other organizations : Jinan University, Chinese University of Hong Kong, Shenzhen, University of Hong Kong, First Affiliated Hospital of Jinan University, Shantou University, Shantou University Medical College, First Affiliated Hospital of Zhengzhou University

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Variable analysis

independent variables

Ki-67 antibody (Dako, Mississauga, ON, Canada)
STAT5A antibody (Proteintech)

dependent variables

Ki-67 proliferation index
STAT5A staining scores

control variables

Paraffin-embedded tumor sections
Antigen retrieval
Blocking with normal serum
Phosphate buffered saline wash
Biotinylated secondary antibody
Peroxidase-conjugated avidin-biotin complex (Dako)
3,3'-diaminobenzidine (Dako) as chromogen
Hematoxylin counterstain

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