Recombinant PEDF Protein Purification

Recombinant rat PEDF (GenBank accession no. NM_177927) was synthesized by Cusabio Biotech, Co., Ltd. (Wuhan, China). In brief, 293T cells were transfected with the 20 μg recombinant vector pGEX 6P-1 (GE Healthcare, Chicago, IL, USA), containing glutathione S-transferase (GST)-tagged PEDF using Lipofectamine^® 3000 according to the manufacturer's protocols. GST-tagged PEDF proteins were purified by high-pressure liquid chromatography purification (>90% purity) and amino-terminal sequence determination (23 ,24 (link)). The resultant proteins were soluble in aqueous solutions.

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Zhao Q., Liu Z., Huang B., Yuan Y., Liu X., Zhang H., Qiu F., Zhang Y., Li Y., Miao H., Dong H, & Zhang Z. (2018). PEDF improves cardiac function in rats subjected to myocardial ischemia/reperfusion injury by inhibiting ROS generation via PEDF-R. International Journal of Molecular Medicine, 41(6), 3243-3252.

Publication 2018

Recombinant rat PEDF pGEX 6P-1 Lipofectamine® 3000

293t cells Glutathione s transferase High pressure liquid chromatography Lipofectamine Pedf Proteins Sequence determination Vector

Corresponding Organization :

Other organizations : Xuzhou Medical College

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Variable analysis

independent variables

Recombinant rat PEDF (GenBank accession no. NM_177927)

dependent variables

Purification of GST-tagged PEDF proteins (>90% purity)
Amino-terminal sequence determination

control variables

293T cells
Recombinant vector pGEX 6P-1 (GE Healthcare, Chicago, IL, USA)
Lipofectamine® 3000 (manufacturer's protocols)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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