Immunohistochemical Evaluation of ARL3 Expression

Immunohistochemical procedures were performed as described previously [28 (link)]. In brief, after deparaffinization, antigen retrieval and endogenous peroxidase activity blocking, sections were blocked with normal goat serum for 15 min and incubated with anti-ARL3 primary antibody (Proteintech, 10961-AP, 1:250) at 4 °C overnight in a humidified chamber. After washing with PBS three times, the sections were incubated with biotinylated goat anti-rabbit IgG (ZSGB-BIO, SP-9001) for 15 min at room temperature. DAB was applied for staining after washing with PBS. The nuclei were counterstained with hematoxylin and the sections were mounted with coverslips after dehydration. Immunohistochemical staining was evaluated with a German immunohistochemical score (GIS) [29 (link)], which further classified the patients into low (GIS < 4) and high (GIS ≥ 4) ARL3 expression groups.

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Wang Y., Zhao W., Liu X., Guan G, & Zhuang M. (2019). ARL3 is downregulated and acts as a prognostic biomarker in glioma. Journal of Translational Medicine, 17, 210.

Publication 2019

SP-9001

Anti antibody Anti igg Antigen Dehydration Goat Hematoxylin Nuclei Patients Peroxidase Rabbit Serum

Corresponding Organization :

Other organizations : First Affiliated Hospital of Shantou University Medical College, Shantou University, Shantou University Medical College, China Medical University, First Hospital of China Medical University

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Variable analysis

independent variables

Anti-ARL3 primary antibody concentration (1:250)

dependent variables

ARL3 expression (classified into low and high groups using German immunohistochemical score)

control variables

Deparaffinization
Antigen retrieval
Endogenous peroxidase activity blocking
Blocking with normal goat serum
Incubation time and temperature of primary antibody
Biotinylated secondary antibody incubation time and temperature
DAB staining
Hematoxylin counterstaining
Mounting with coverslips

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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