Immunoblotting of Cellular Protein Markers

Lysates were prepared from fresh tissues and cells and then subjected to immunoblotting (IB) in accordance with a previously described protocol (Liu et al., 2017 (link)). Proteins were then detected using a range of antibodies: mouse anti-GFAP (1:2000, Proteintech, Chicago, IL, United States), mouse anti-NEUN (1:500, Millipore, Belecula, CA, United States), rabbit anti-BAF45D (1:500, Proteintech, Chicago, IL, United States), mouse anti-beta-III-tubulin (1:1000, Arigo) and rabbit anti-GAPDH (1:2000, Proteintech, Chicago, IL, United States). Specific details of the antibodies used for IF and IB are given in Supplementary Table S2.

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Wang Z., Huang J., Liu C., Liu L., Shen Y., Shen C, & Liu C. (2019). BAF45D Downregulation in Spinal Cord Ependymal Cells Following Spinal Cord Injury in Adult Rats and Its Potential Role in the Development of Neuronal Lesions. Frontiers in Neuroscience, 13, 1151.

Publication 2019

mouse anti-GFAP mouse anti-NEUN rabbit anti-BAF45D mouse anti-beta-III-tubulin rabbit anti-GAPDH

Antibodies Beta tubulin Cells Gapdh Gfap Mouse Proteins Rabbit Tissues

Corresponding Organization : First Affiliated Hospital of Anhui Medical University

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Variable analysis

independent variables

Preparation of lysates from fresh tissues and cells

dependent variables

Immunoblotting (IB) for protein detection

control variables

Previously described protocol (Liu et al., 2017)
Antibody concentrations: mouse anti-GFAP (1:2000), mouse anti-NEUN (1:500), rabbit anti-BAF45D (1:500), mouse anti-beta-III-tubulin (1:1000), rabbit anti-GAPDH (1:2000)

controls

Positive controls not explicitly mentioned
Negative controls not explicitly mentioned

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