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In the first protocol, crude plasma samples were subjected to tryptic cleavage (Series #1).
Compared with the first series of experiments, we were able to reliably detect four proteins not previously detected in crude plasma, including von Willebrand factor, which was detected in one of the samples in a single technical run.
In a series using crude plasma five proteins remained undetected, while application of clean-up and depletion strategies reduced the number of undetected proteins to one and zero, respectively.
The resulting lysate was further fractionated into crude plasma membrane and cytoplasmic fractions by centrifugation at 100,000g for 1 hr at 4°C.
Acomparison of the traditional biotin-streptavidin system with the sucrose gradient-based extraction kit shows that the sucrose gradient-based extraction method yields a highly enriched, pure, plasma membrane fraction, while the biotin-streptavidin method produces a crude plasma membrane fraction with considerable contamination of cytoskeletal proteins.
Label-free protein quantification was performed in crude plasma only.
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In contrast, crude plasma-induced breakage and roughness was shown in the cell surfaces after a 120 min treatment.
Because the NanoString method does not require polymerase activity, says Beechem, it works in less-than-ideal conditions like crude lysates, plasma, or formalin-fixed paraffin-embedded (FFPE) samples, which is how clinical tissue specimens are often stored.
The main advantage of our ELISA design is its ability to correctly estimate the neutralization capacity of crude hyperimmune plasma and antivenom sera independently of their antibody composition in terms of whole IgG or F ab′ 2 fragment.
Effect of crude crocodile plasma on cell viability was tested by MTT assay.
The effect of the crude crocodile plasma on the cell of Salmonella typhi ATCC 11778 tested as a reference strain was examined by SEM.
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