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Proteins, Expression, Isolation and Analysis | … Eluates with >90% purity were dialyzed against Tris buffer. The endotoxin level in PEDF was controlled using an endotoxin assay kit as described by the manufacturer (Genscript Biotech Corp, Nanjing, China). 2.6. Co-immunoprecipitation … | Get A Quote |
Pro-inflammatory cytokines, such as tumor necrosis factor alpha (TNFα), may contribute to hepatic steatosis in the situation of excess lipid accumulation in farmed fish. Pigment epithelium-derived factor (PEDF) is an endogenous anti-inflammatory factor and promotes lipolysis. Accordingly, we isolated PEDF from grass carp and investigated its role in TNFα-induced hepatic steatosis. Sequence analysis showed that PEDF gene, which possesses 8 exons and 7 introns, encodes a protein with 409 amino acids. PEDF was a critical determinant of the transcriptional response to nutrient availability in grass carp. Endogenous PEDF was an intracellular protein with cytoplasmic distribution and directly interacts wi... More