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Briefly, an affinity purified sheep polyclonal anti-hTERT antibody (20 µg) was added per ml of cleared lysate from HEK-293T cells and rotated for 1 h at 4 °C.
Nuclei and cellular debris were pelleted by centrifugation at 1000 rpm for 8 min. Then, 1 mL of cleared supernatant was mixed with 1 mL of 85% sucrose in TNEV and transferred to the bottom of a Beckman 14×95-mm 14×95-mmge tube.
1.5 ml of GSH bead slurry (GE Healthcare) was added to 50 ml of cleared lysate.
The HaloLink resin (1.0 ml) was added to 10 ml of cleared lysate.
Antibody-coupled beads were washed and equilibrated with lysis buffer before incubating with 0.5 ml of cleared cell lysates for 16 hr at 4°C.
Untransfected cells were plated onto round coverslips and incubated for 24 hr in 2 ml of cleared CLCA1- or pHLsec-conditioned medium supernatants.
Similar(51)
A 2.5-ml aliquot of cleared culture supernatants from replicon cells was added to approximately 70% confluent of Huh-7 cells in 25-cm flasks, and the same amount of complete DMEM was added 2 h later.
Once 500 mL of clear fluid was tolerated, they received a regular diet.
Take 5 ml of clear filtrate.
Over 20 ml of clear ascitic fluid was immediately drained, and compression of the tense abdomen was released.
A 14-French silicone Foley catheter was inserted per urethra without any delay and 300 ml of clear urine was drained.
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CEO of Professional Science Editing for Scientists @ prosciediting.com