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Mixed reagent was prepared by adding 1 mL of Reactive 1 (1% Potassium sodium tartrate) to 50 mL of Reactive 2 (2 g of Sodium carbonate) in 100 mL of 0.1 M NaOH.
The protein solutions were mixed with 0.5 mL of reactive Alexa Flor 488 and Texas Red dyes, respectively, for 1 h at room temperature.
0.5 mL aliquot of the OC extract was transferred to a glass tube containing 0.5 mL of reactive FC, after 5 min 2 mL of Na2CO3 solution (200 mg/mL) was added.
To the hemolysate or 0.5 mL of plasma, 4.5 mL of reactive mixture containing 0.375% of TBA and 15% of trichloroacetic acid (TCA) in 0.25 M HCl was added.
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20 ml of each reactive antiserum were purified by binding to ∼5 mg of the respective recombinant protein coupled to CNBr-activated sepharose beads (Amersham), washed extensively with PBS and elution with first 0.2 M glycine, pH 3, then 0.2 M triethanolamine, pH 9. Affinity-purified polyclonal antibodies were dialysed against PBS and concentrated by ultrafiltration (Amicon Ultra 50k, Millipore).
(5 mL) of cold reactive dye into a cup.
(1) Enzyme Activity (U / mL / min ) = Sugar released µ g × Total volume of reactive media mL × Dilution factor (DF ) Molecular weight of maltose × Enzyme used mL × Time of incubation (min ).
Kidney samples were weighed and digested in a solution (2 1) of nitric acid (9601 02, Reactive Grade, J.T. Baker, 69.9% of purity) and hydrochloric acid (9535 02, Reactive Grade, J.T. Baker, 37.1% of purity) plus 3 mL of hydrogen peroxide (2186, Reactive Grade, J.T. Baker, 30% of purity) overnight.
These reduced thiolated DNA strands (1 μM) were then reacted with 1 mL of the functionalized (thiol-reactive) BTO NC solution to obtain 1 mL of DNA BTO conjugate solution (at ∼60 °C for 30 min to 1 h).
Taddei ML, Parri M, Mello T, Catalano A, Levine AD, Raugei G et al. Integrin-mediated cell adhesion and spreading engage different sources of reactive oxygen species.
Strawson's theory of "reactive attitudes".
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