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The cultures were centrifuged at 18000 × g for 10 min to collect the bacteria, the pellets were dissolved in SDS-PAGE sample buffer, and the supernatants were separated out and then filtered through a 0.22-μm low protein binding filter.
Supernatant was filtered (0.45 μm low protein binding filter; Sartorius Stedim Biotech) and used immediately for transduction of Neuro2A cells, or concentrated for in vivo administration by centrifugation at 20 000 g (4 h at 4 °C) and overnight resuspension in 1.5 ml phosphate-buffered saline (PBS; 4 °C).
The supernatant containing whole-cell protein extracts was filtered through a 0.22 µm-pore-size low protein binding filter.
Digested gluten or bread samples were centrifuged for 10 min at 13,400g and filtered through a 0.2 µm low protein binding filter.
To further characterize nucleic acid binding, filter binding assays were performed by preloading Gadd45a with unlabeled nucleic acids followed by competition with labeled RNA from a plasmid multiple cloning site (Figure 1B, C).
Supernatant were pooled, centrifuged to remove cells and then filtered through 0.45 μm low protein binding filter.
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"I'm just kind of unimpressed with what they're announcing," says Brown. "They're coming to market so late, with requirements that were established years and years ago". For example, the new Google Search Appliance can search up to 1 billion documents and includes security features like early binding filtering that can skew results based on what the user needs.
For example, the new Google Search Appliance can search up to 1 billion documents and includes security features like early binding filtering that can skew results based on what the user needs.
The RNA binding of the purified NP was examined by a filter binding assay employing both protein and nucleic acid binding filters (data not shown).
We previously described an efficient method for the collection of relatively large quantities of Culicoides saliva in a pure form, using protein binding filters [27].
Culture filtrate proteins (CFP) were collected by passing spent broth medium through 0.25 µm low-protein binding filters (Corning, Lowell, MA) and concentrating the eluent using Amicon Ultracell-15 (10,000 molecular weight cutoff) spin columns.
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