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Western blot analyses of brain samples were performed as described in Beaulieu et al. [28].
Lipidomic analyses of brain samples were performed with a microfluidics ionKey/MS system composed of an ACQUITY UPLC M-Class, the ionKey source, and an iKey CSH C18 130 Å, 1.7 μm particle size, 150 μm × 100 mm column (Waters Corporation, Milford, Massachusetts, USA) coupled to a Synapt G2-Si (Waters Corporation, Manchester, UK).
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Proteinase K digestion of brain samples was performed as described [27].
The qRT-PCR analysis of human brain samples was performed using our established methods [55].
Western blot analysis from adult brain samples was performed as described by Porta et al. with minor modifications [5].
Storage and biochemical analysis of human brain samples was performed with consent from relatives and with approval from the Local Research Ethics Committee of the Institute of Neurology/National Hospital for Neurology and Neurosurgery (London, U.K).
ChIP from brain samples was performed using 50 70 mg of tissue homogenized using a tissue grinder after crosslinking and quenching, and following the same procedure used for fibroblasts with the exception that samples were incubated overnight at 4 °C with 2 μg of anti- pan H3 anti- pan H3Millipore), andwell as anti-H4K9mEMD H3K27Millipore9me3, H4K20me3 (Diasenode).
All samples were performed in duplicates.
All samples were performed in triplicate.
Chromatography of brain microdialysate samples was performed on a Beckman Coulter™ Ultrasphere® 5 μm C-18 column (2 mm I.D. × 250 mm, Alltech) at a constant temperature of 30°C.
Extraction of blood and brain samples was then performed by a method based on protein precipitation using acetonitrile containing a structural analogue of -U50,488 as an internal standard.
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