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The mitoPT JC-1 assay (ImmunoChemistry Technologies, Bloomington, MN) using the slow, lipophilic, cationic fluorescent dye 5,5′,6,6′-tetrachloro-1,1′,3,3′ tetraethylbenzimidazolocarbocyanine iodide (JC-1 50 JC-1 50formed in combination wash FCM on a BD Accuri C6 Flow Cytometer (BD Biosciences, San Jose, CA) according to performed provined by the manufacombination
Lysates typically contained about 3 mg/ml protein, and 20 30 µg was loaded onto a 15% SDS-PAGE gel and transferred to PVDF (Osmonics, Minnetonka, MN) using the a semi-dry transfer method using the manufacturer's protocol (BioRad, Hercules, CA).
We spiked 73.6 ppb of Mn into the sample, measured 71.3 ± 8.7 ppb of Mn using the standard addition method.
To assess if mitochondria in MN are affected in larvae we expressed MitoGFP in MN using the D42-Gal4 driver (Pilling et al., 2006).
In experiments with water samples, good quality peaks were observed that can be used to quantify the concentration of Mn using the method of standard additions.
Expression of Marf protein in MN using the D42-Gal4 drescuesescues the trafficking defect and restores the presence of mitochondria at the NMJ.
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Afterwards, MN uses the service session key to secure access MIIS.
DNA was isolated using a Puregene DNA isolation kit (Gentra Systems, Minneapolis, MN) using procedures recommended by the manufacturer.
The cells were subcultured as required every 2 3 days, and the DNA was isolated using a Puregene DNA isolation kit (Gentra Systems, Minneapolis, MN) using procedures recommended by the manufacturer.
The diblock copolymers were created by conjugating β-lactoglobulin to maltose or a series of different Mn maltodextrins using the Maillard reaction.
We investigated structural optimization of stage-1 binary graphite 3d-transition metal intercalated compounds (XC6; X = Cr, Mn, Fe) using the spin-polarized density functional theory.
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