Tissue Array and IHC Analysis in Breast Cancer

Tissue Array Preparation: We followed the methods described by Wang et al.^{28 (link)}. In brief, the Quick-Ray^® UT-06 (Unitma Co., Ltd., Seoul, Korea) tissue microarray system and the Quick-Ray premade recipient block (UB-06) wax model were used to prepare tissue specimens (1 mm in diameter). Two representative sites from each breast cancer tissue sample were selected for sampling. IHC Analysis: IHC staining of paraffin-embedded tissue sections used the Envision System (Dako, Glostrup, Denmark), as described previously^{25 (link),26 (link)}. Primary antibodies consisted of anti-WTAP rabbit monoclonal antibody (clone EPR18744, diluted at 1:3200; Abcam, Cambridge, England), ready-to-use anti-ER rabbit monoclonal antibody (clone SP1, Dako), ready-to-use anti-PR mouse monoclonal antibody (clone PgR636, Dako), ready-to-use anti-Podoplanin mouse monoclonal antibody (clone D2-40, Dako) and HercepTest (Dako). The secondary antibody was Dako’s HRP rabbit/mouse universal antibody (Dako, Glostrup, Denmark). The negative control was incubated with vehicle then with secondary antibody, without primary antibody.

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Wang C.Q., Tang C.H., Wang Y., Huang B.F., Hu G.N., Wang Q, & Shao J.K. (2022). Upregulated WTAP expression appears to both promote breast cancer growth and inhibit lymph node metastasis. Scientific Reports, 12, 1023.

Publication 2022

Envision System anti-WTAP rabbit monoclonal antibody (clone EPR18744, ready-to-use anti-ER rabbit monoclonal antibody (clone SP1, ready-to-use anti-PR mouse monoclonal antibody (clone PgR636, HercepTest HRP rabbit/mouse universal antibody

Antibodies Antibody Breast cancer Clone Embedded paraffin Microarray Monoclonal antibody Mouse Rabbit Staining tissue Tissue

Corresponding Organization : Dongyang People's Hospital

Other organizations : China Medical University

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Variable analysis

independent variables

Tissue microarray system used (Quick-Ray® UT-06)
Premade recipient block (UB-06) wax model used
Primary antibodies used (anti-WTAP, anti-ER, anti-PR, anti-Podoplanin, HercepTest)

dependent variables

IHC staining results of paraffin-embedded tissue sections

control variables

Tissue specimen size (1 mm in diameter)
Number of representative sites sampled from each breast cancer tissue sample (2)

positive controls

Not specified

negative controls

Incubation with vehicle then with secondary antibody, without primary antibody

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