Tissue Microarray-based IHC Profiling

We used tissue microarrays (F048Br01a, Bioaitech, China) comprising 24 BC tissues of varying stages and grades, along with their corresponding adjacent tumor tissues. IHC staining and scoring were conducted following a previously described protocol [20 (link)]. In summary, following the removal of paraffin, rehydration, and microwave antigen retrieval, the slides were left to incubate with antibodies overnight at a temperature of 4 degrees Celsius. Anti-ERBB2 (ab237715, Abcam), anti-SDC1 (ab130405, Abcam), and anti-MMP14 (ab51074, Abcam) were each diluted to concentrations of 1:2000, 1:1000, and 1:800, respectively. Afterward, the slides were subjected to secondary antibodies at ambient temperature for a duration of 30 minutes. Then, they were stained using DAB substrate and subsequently counterstained with hematoxylin.

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Fang Y., Zhang Q., Chen Z., Guo C, & Wu J. (2024). Clinical significance and immune characteristics analysis of miR-221-3p and its key target genes related to epithelial-mesenchymal transition in breast cancer. Aging (Albany NY), 16(1), 322-347.

Publication 2024

ab237715 ab130405 ab51074

Corresponding Organization :

Other organizations : Shantou University, Cancer Hospital of Shantou University Medical College, Jiangmen Central Hospital

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Variable analysis

independent variables

Tissue microarray (F048Br01a, Bioaitech, China) comprising 24 BC tissues of varying stages and grades
Adjacent tumor tissues

dependent variables

ERBB2 expression
SDC1 expression
MMP14 expression

control variables

Removal of paraffin
Rehydration
Microwave antigen retrieval
Incubation with antibodies overnight at 4 degrees Celsius
Concentrations of antibodies: ERBB2 (1:2000), SDC1 (1:1000), MMP14 (1:800)
Incubation with secondary antibodies at ambient temperature for 30 minutes
DAB substrate staining
Hematoxylin counterstaining

controls

No positive or negative controls were explicitly mentioned in the provided information.

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