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The catalytic parameters of individual kinase variants were measured using a spectrophotometric coupled-enzyme assay.
The basal autophosphorylation rates of CheA variants were measured as previously described.
Association and dissociation rates of IL-4 variants to immobilized IL4-RαECD variants were measured on a BIA2000 system.
Arabidopsis plants were exposed to 4°C, and the levels of alternatively spliced RNA variants were measured by qRT-PCR.
The kinetics of solubilization of DMPC multilamellar vesicles (MLV) by the apoE variants were measured by monitoring the decrease in absorbance at 325 nm, as described previously.
The activities for each of these P450 variants were measured on all six substrates (12-pNCA, 2PE, PROP, 11PA, 2A5C, and MDOB).
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Expression of the different hPolλ protein variants was measured by immunoblotting of total cellular extracts (75 µg) with Polλ polyclonal antibodies [4] and the actin antibodies AC-40 Sigma-Aldrich Chemical Coo). Sigma-Aldrich Chemical Co
The stability of the IGPS clamp-deletion variants was measured by urea denaturation as described previously [17] in a buffer containing 10 mM potassium phosphate, pH 7.8 for sIGPS and pH 7.0 for eIGPS, 0.2 mM K2EDTA, and 1 mM βME.
Thus, the kinetic solubility of a selection of active variants was measured at 37 °C, pH 7.0 (Fig. 5).
Furthermore, many previous studies have not adequately defined the precise ER β variants being measured or have used antibodies capable of detecting multiple variants (Skliris et al, 2003).
In the second approach, successive and accumulative single random substitutions were introduced, and the k-tuple distance between each pair of new and previous variants was measured.
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