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HPLC charts of recombinant mouse αsyn used in this study.
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Alternatively, βSyn could be fully purified from fractions containing αSyn using HIC.
The expression of human αSyn using AAV vectors was used to specifically induce pathological changes in the rat nigrostriatal system.
The discrepancy may have arisen from differences in the test conditions, because in the previous study, we used female mice injected with human αsyn fibrils, whereas in this study we used male mice injected with mouse αsyn.
Mouse αsyn cDNA in bacterial expression plasmid pRK172 was used.
Pre-embedding immunoelectron microscopy (IEM) was used then to define the subcellular localization of both transgenic (human) and endogenous (mouse) αSYN in the brains of transgenic MSA mice 12 weeks after treatment with PSI or vehicle.
For oral administration, 2- or 3-month-old C57BL/6 J mice were orally administrated with 400 μg of human αsyn monomer, human αsyn fibrils, mouse αsyn monomer or mouse αsyn fibrils every two weeks for 4 times.
We investigated the spread of αsyn pathology in brains of mice after unilateral injection of recombinant mouse αsyn fibrils into SN, Str, or EC.
Mouse αSyn from a commercially available mouse brain extract (Sigma) migrated slightly higher than the yeast-expressed human αSyn, possibly due to the seven-amino-acid differences between the two proteins.
Rabbit polyclonal pS396 antibody (Calbiochem) is specific for phosphorylated tau at serine 396; biotin-AT8 (Thermo Scientific) is specific for phosphorylated tau at serine 202/threonine 205; anti-mouse αsyn rabbit monoclonal antibody (Cell Signaling Technology) is specific for mouse αsyn.
For intraperitoneal injection, 2-month-old C57BL/6 J mice were injected intraperitoneally with 100 μg of mouse αsyn monomer or fibrils.
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