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All the reactions were performed in triplicate in 96-well micro plates and monitored in a Spectra Max 340 (Molecular Devices, USA) spectrometer.
The cell suspensions were then transferred into 100-well sterile micro plates and incubated at 37°C without shaking.
All samples were measured in duplicate (50 μl) within a standard range of 0-3000 ng/ml and with a total incubation time of 2 hours and 10 minutes, in room temperature, using two monoclonal antibodies, one in the solid phase (coated micro plates) and the other in the conjugate buffer.
For quantification of NST-732 uptake, homogenized tumor samples were aliquoted (in triplicates) onto black Ritter flat bottom micro plates, and read at 360 nm excitation and 520 nm emission, using fluorescence micro plate reader (GENious Fl Reader, Tecan, Grodig, Austria).
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The peptide containing supernatant was transferred into a new micro plate and the extraction was repeated.
Add 100 μl of endotoxin test solution, endotoxin standard solution and samples into each hole of the micro plate and placed it tachypleus amebocyte lysate, 37°C for 10 minutes.
1C11 cells were cultured on glass cover slips at the bottom of 24 wells-micro plates and induced to differentiate into 1C115-HT cells.
From a detailed variational procedure the governing equilibrium equation of the micro-plate and the most general boundary conditions are derived, in terms of the deflection, using the principle of minimum potential energy.
Details of preparation of Ag coated Micro-plates and procedure to test the sera.
Aliquots (200μl) were transferred to a 96 well flat-bottomed micro-plate and read on a plate reader (Thermo Multiskan spectrum) at 510 nm.
The resultant mixture (160 μL) was loaded onto a flat bottom-micro plate and the optical density read at 562 nm.
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