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Fragments were generated to a variety of sizes, but all of them were smaller than 10 kDa when resolved by size exclusion chromatography (Fig. 1A) or SDS-PAGE (Fig. 1B; lane 6).
3) The mass of the complexes resolved by size exclusion chromatography has been assessed by multi-angle lights scattering (MALS, a method orthogonal to migration through a gel filtration matrix) (revised figure 2-supplement 1).
The bacterially-expressed proteins were purified by affinity tag (His 6X) chromatography, phosphorylated to completion by the kinase PERK, in vitro and resolved by size exclusion chromatography on a Superdex 75 30/300 mm column.
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The recombinant protein was eluted with 500 mM imidazole and was further resolved by size-exclusion gel chromatography (SEC, Superdex 75 26/60; GE Biosciences, Piscataway, NJ).
(A ) UV protein absorbance trace of a PPP1R15B 630 701 -PP1G(7–300) binary complex expressed in bacteria and resolved by size-exclusion chromatography.
(A ) UV protein absorbance trace of a PPP1R15A (539–614)-PP1G(7 323 -G-actin 7 323 -G-actin 7 323 -G-actin from the banderially-expresseDNaseary complex, rabbIt muscomplexctin assemblede pancreatic DNase I, resolved by size-exclusion chromatography.
(A ) UV protein absorbance trace of a PPP1R15B 631 701 -PP1G(7 323 -G-actin 7 323 -G-actin 7 323 -G-actincomplexly-expressed binassembledex and rabbit muscle G-actin and resolved by size-exclusion chromatography.
(A ) UV protein absorbance trace of a PPP1R15B-PP1G co-expressedxpressed in bacteria as a GST-PPP1R15B 631 701 -MBP GST-PPP1R15B 631 701 -MBPuntaGST-PPP1R15B 631 701 -MBPd by glutathione afusiony chromatograprotein, alongsideavage of the GST tag, resolved by size-excluntaggedromatograPP1G
UV protein absorbance trace of a PPP1R15A 539 614 -PP1G(7 323 -G-actin 7 323 -G-actin 7 323 -G-actincomplexly-expressed binassembledex and rabbit muscle G-actin and resolved by size-exclusion chromatography on tandem S200 and S75 30/300 columns.
Ties were resolved by ecoregion size for species richness (the smaller the size the higher the rank).
The objective of the current research targets mainly on the accurate capturing of Navier Stokes diffusive interfaces, where the thickness can be resolved by the cell size.
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