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Proteinase K (PK) digestion and membrane protein analyses were performed as described (Zhao et al, 2009).
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The availability of pure, stable and well-behaved protein is the key factor for successful structural studies for all membrane protein structural analyses.
Since EphA4 is an integral membrane protein we analysed whole tissue lysates (containing soluble and insoluble fraction).
In addition, the taxonomic distribution of the identified outer membrane proteins was analysed to illuminate their conservation throughout the chlamydiae and among representatives of all other bacterial lineages.
To establish whether this process reflected a specialized pathway or a more general feature of membrane protein biogenesis, we analysed an even simpler potassium channel subunit with only two TMs separated by a putative pore loop, the viral Kcv protein [ 28].
These recent studies build on classic studies of protein secretion in bacteria, of membrane protein crystallization and biophysical analyses.
Soluble and membrane protein fractions were then analysed by SDS-PAGE and subsequent Western blotting.
Analysing the structure of membranes and membrane proteins remains a challenging problem, given the difficulty associated with purifying membrane fractions and analysing membrane protein structures (Elofsson & von Heijne, 2007).
The trifluoromethyl phenylalanine (tfmF) was successfully used for analyses of membrane protein's conformation changes, dynamics and functions (Shi et al., 2012).
As would be expected for integral membrane protein, SDS-PAGE and immunoblot analyses of fractionated lysates from phmt-1-hmt-1 GFP phmt-1-hmt-1 GFPHMT-1::GFphmt-1-hmt-1 GFP phmt-1-hmt-1 GFP, but not among soluble proteins (Fig. 1A).
Efficient reconstitution of membrane proteins for functional analyses can be achieved by dilution of a ternary mixture containing proteins, lipids and detergents.
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