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The successfully received packets will be stored into the circular buffer with the corresponding flag being set true.
To avoid fabrication-related variation of the hybridization signals the hybridization assays were performed on the same chip, initially with RNA and subsequently, after regeneration of the microarray (by heating to 70°C in pure hybridization buffer), with the corresponding DNA targets.
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The resulting clarified homogenates from the various tissues were loaded onto multistep sucrose gradients (1.08, 1.13, 1.18 and 1.26 g/cm) buffered with the corresponding homogenization buffer.
Aliquots of the lysate were then adjusted to a final working concentration of 0.7 μg/μl using the assay buffer provided with the corresponding ELISA kit.
Experiments were performed by incubating the cells with the saline buffer complemented with the corresponding indole (analytical standard grade, Sigma Aldrich) amount diluted from a 500 mM stock in ethanol.
They were then imaged in HBSS buffer supplemented with the corresponding peptide at RT. Epifluorescence imaging was performed on a Zeiss Axiovert 200 M Microscope equipped with a xenon lamp and a cooled CCD, under a 40X oil immersion objective.
They were thereafter incubated overnight at 4°C with the rabbit anti-MMP-1 (1/500) primary antibody diluted in blocking buffer and then with the corresponding peroxidase-conjugated secondary antibody for 1 h at room temperature.
Then, cells were centrifuged and washed four times with FACS buffer before incubation with the corresponding FITC-conjugated secondary antibody for 30 min. Then, the cells were centrifuged and washed four times in FACS buffer, and the cell surface CD44 expression level was measured by flow cytometry (BD Accuri C6 flow cytometer).
After washing three times in Tris-buffered saline Tween buffer, nitrocellulose membranes were incubated with the corresponding secondary antibody (Alexa Fluor 647 goat anti-mouse IgG antibody or Alexa Fluor 647 goat anti-rabbit IgG, 1 2,000; Molecular Probes, Darmstadt, Germany).
After washing the beads with the corresponding buffer, bound proteins were eluted with buffer lacking Ca2+ ions.
If args are saved (no explicit userid in the submit call), they are stored locally and become the active buffer on the corresponding working return.
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