Tissue Microarray Proteome Extraction

All tissue microarray chips were purchased from Shanghai Outdo Biotech Co. Ltd. Tumor and paracancerous tissues (normal) were freshly excised from a patient with NSCLC undergoing surgery. Tumor tissue and matched paracancerous tissue were homogenized (53 (link)). Briefly, the specimens were cut into 0.5-mm sections before digestion with 0.1% collagenase IV (Gibco, no. 17104019) for 1 hour at 37°C. The cells were then passed through a 70-μm cell strainer (BD, no. 352350) and collected by centrifugation for 15 min at 400g. Plasma membrane proteome extracts were prepared from single-cell suspensions of tissues.

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Wang Z., Li Y., Hou B., Pronobis M.I., Wang M., Wang Y., Cheng G., Weng W., Wang Y., Tang Y., Xu X., Pan R., Lin F., Wang N., Chen Z., Wang S., Ma L.Z., Li Y., Huang D., Jiang L., Wang Z., Zeng W., Zhang Y., Du X., Lin Y., Li Z., Xia Q., Geng J., Dai H., Yu Y., Zhao X.D., Yuan Z., Yan J., Nie Q., Zhang X., Wang K., Chen F., Zhang Q., Zhu Y., Zheng S., Poss K.D., Tao S.C, & Meng X. (2020). An array of 60,000 antibodies for proteome-scale antibody generation and target discovery. Science Advances, 6(11), eaax2271.

Publication 2020

tissue microarray chips collagenase IV

Cell Centrifugation Chips Collagenase 1 Digestion Microarray Nsclc Patient Plasma membrane Proteome Surgery Tissues Tumor

Corresponding Organization : State Key Laboratory of Oncogene and Related Genes

Other organizations : Duke Medical Center, Duke University Hospital, Duke University, Microbiology Institute of Shaanxi, Chinese Academy of Sciences, Guangxi Academy of Agricultural Science, China Agricultural University, Zhengzhou Fruit Research Institute, Chinese Academy of Agricultural Sciences, Institute of Crop Sciences, Southwest University, China-Japan Friendship Hospital, Karolinska Institutet, Ludwig Cancer Research, South China Agricultural University, Wuhan University

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Variable analysis

independent variables

Tissue type (tumor tissue vs. paracancerous tissue)

dependent variables

Plasma membrane proteome extracts

control variables

Patient with NSCLC undergoing surgery
Sample preparation (cutting into 0.5-mm sections, digestion with 0.1% collagenase IV for 1 hour at 37°C, passing through a 70-μm cell strainer, centrifugation for 15 min at 400g)

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