Identification of GRP78 and vaspin complex by tandem affinity tag purification. Identification of GRP78 and vaspin complex by tandem affinity tag purification.

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Identification of GRP78 and vaspin complex by tandem affinity tag purification. Identification of GRP78 and vaspin complex by tandem affinity tag purification. A: pCTAP-vaspin-Ad was applied to HepG2, and purified products were analyzed by SDS-PAGE. In-gel digestion with trypsin and analysis by LC-MS/MS revealed that band 1 is GRP78 and band 2 is vaspin. B: HepG2 was treated with vaspin recombinant protein. Vaspin-GRP78 complex was immunoprecipitated by vaspin antibody and blotted with GRP78 antibodies. C: Pull-down assay using GST-vaspin fusion protein and in vitro–translated GRP78. D: Pull-down assay using GST-GRP78 fusion protein and in vitro–translated vaspin. (A high-quality color representation of this figure is available in the online issue.)‏ Atsuko Nakatsuka et al. Diabetes 2012;61:2823-2832 ©2012 by American Diabetes Association