Exact(8)
After 10 min on ice, the samples were spun at 16,000× g for 10 min. Both supernatant (i.e,ytosol) and pellet (i.e., membrane) fractions were collected and placed in fresh microcentrifuge tubes.
The standard water immunomagnetic separation (IMS) procedure (GC-Combo; Dynal Biotech, Invitrogen) was conducted in duplicate on 10 ml volumes each containing not more than 0.5 ml pellet (i.e. a maximum of 5% of the filter was analysed).
Now, like a mouse that's found an experimental lever that only sometimes gives up a pellet, I find myself repeatedly pressing it, hoping to hit on the magical combination that will get my bank to behave as though security was something that a reasonable, sane person could understand, as opposed to a magic property that arises spontaneously in the presence of sufficient obfuscation and bureaucracy.
The pellet (Pellet I) was used to prepare the nuclear fraction.
"Pellet I" (∼9 g) was harvested by centrifugation at 9000 g.
Three g of pellet I was suspended in PBS and lysed by tip sonication in an ice bath.
Similar(52)
Meanwhile, the cell monolayers were incubated with trypsin, which detaches the cells and removes surface bound TF, and centrifuged to separate the surface bound (supernatant) and intracellular (pellet) I-rhTF.
In the pharmaceutical industry, the coating process is often applied on pellets, i.e., small spherical particles with a narrow size distribution, which are then enclosed in capsules or compressed into tablets and contain the active pharmaceutical ingredient.
Regarding the acidity measurements, it must be stressed that any effects of the supported metals on these parameters would be limited to the "shell" region of the catalyst pellets, i.e., the "core" region acid base properties would not be affected.
Samples of the supernatants and pellets from the various steps in this procedure were then immunoblotted and β1 integrin, actin, transferrin receptor, and EGFR were all detected in the conditioned medium from NeoA-treated cells and in the pellets (i.e. the vesicle-containing fraction) of the samples from the NeoA-treated, but not from the vehicle control, DMSO-treated, cells (Figure 6C).
RNA was prepared from the frozen bacterial pellets using modifications to the Promega SV total RNA isolation kit protocol, published elsewhere [2] except that double the initial volumes were used to take into account the larger bacterial pellets i.e. 200µl starting volume.
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