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The solution of peptide was diffused and distributed evenly on the device due to the surface tension.
The stock solution of peptide standards was prepared in a 1 mM concentration, and working standard solutions of 25 pM were prepared through the dilution of stock solution.
Control groups received either a mixture of 10 µg free doxorubicin and peptide RMS-P3/RR, an equimolar solution of peptide RMS-P-3/RR alone or PBS alone.
Beads in each aliquot were incubated for 1 hour at room temperature with 200 µl of a 200 µg/ml solution of peptide C12 (a mixture of 160 µl of the solubilization buffer S2 and 40 µl of the stock aqueous solution (1 mg/ml) of C12 peptide) on a rocking platform.
The initial protein concentration was 92 μM and volumes of 2 mM stock solution of peptide were added until the protein:peptide ratio was 1 4.
Cysteine-terminated peptides were immobilized onto 1% maleimide-presenting SAMs by immersing the monolayers in a solution of peptide (1 mM in TRIS buffer, pH = 7.5) and incubated at 37 °C for 1 h.
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Standards for spectral calibration consisted of a mixed solution of peptides ranging between m/z 900 to 3500 Da.
A 200 μM solution of peptides in PBS/Tween 20 were filled in wells of polypropylene microtitre plates (Greiner Bio-One).
A solution of peptides was filtrated via Millipore (Millipore ZTC18M096) and mixed with the same volume of a matrix solution consisting of saturated a-cyano-4-hydroxycinnamic acid (CHCA) in 50 % ACN containing 0.1 % TFA.
Our laboratory has recently developed liquid crystalline solutions of peptide amphiphile nanofibers that form aligned domains at exceedingly low concentrations (<1wt%), and can be trapped as gels with macroscopic alignment using low shear rates and ionic crosslinking.
While a 200 µM-solution of peptide 131 151 remained highly soluble in water, phosphate-buffered saline (PBS) and culture medium (Figure 2C), aggregates form in the P140 solution at concentrations equal or superior to 50 µM in culture medium and PBS.
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