Immunohistochemical Staining of Akt in Xenograft Tumors

The IHC staining was performed on cryostat sections (4 μm/section) of 786-O xenograft tumors, and the protocol was described in detail in previous studies [12 (link),25 (link)]. The primary antibody (anti-Akt Ser-473, 1: 50, Cellular Signaling Tech), and horseradish peroxidase (HRP)-coupled secondary antibody (Santa Cruz) were applied [12 (link)]. The positive staining was performed through peroxidase activity using the 3-amino-9-ethyl-carbazol (AEC) method (Merck, Shanghai, China).

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Pan X.D., Gu D.H., Mao J.H., Zhu H., Chen X., Zheng B, & Shan Y. (2017). Concurrent inhibition of mTORC1 and mTORC2 by WYE-687 inhibits renal cell carcinoma cell growth in vitro and in vivo. PLoS ONE, 12(3), e0172555.

Publication 2017

horseradish peroxidase (HRP)-coupled secondary antibody

Antibody Horseradish peroxidase Peroxidase Tumors Xenograft

Corresponding Organization : Affiliated Hospital of Nantong University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Akt Ser-473 protein expression (measured through IHC staining)

control variables

786-O xenograft tumor sections
Cryostat sections (4 μm/section)
Primary antibody (anti-Akt Ser-473, 1:50, Cellular Signaling Tech)
HRP-coupled secondary antibody (Santa Cruz)
Peroxidase activity using the 3-amino-9-ethyl-carbazol (AEC) method (Merck, Shanghai, China)

controls

Positive control: none mentioned
Negative control: none mentioned

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