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Protein A-purified neutralizing rabbit anti-HMGB1 IgG (50 µg/ml) was produced and also used for neutralization assays.
Both the virus and the reagent used for neutralization influence the assay sensitivity.
The results showed that the degree of correlation between the two assays was dependent on the reagents used for neutralization.
Both the virus and the reagent used for neutralization (here monoclonal antibodies and sCD4) contribute to the outcome of assay sensitivity, which in certain combinations is also influenced by the target cell used.
Viral RNA was extracted from virus culture supernatant (the same aliquot used for neutralization) as described by Boom et al, [42] followed by reverse transcription using Oligo dT primer (Promega Corporation, Madison, WI) and AMV reverse transcriptase (Access RT-PCR system; Promega Corporation, Madison, WI).
Aqueous ammonia (Merck) was used for neutralization of the HPLC fractions.
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It has been hypothesized that the lower CCR5 concentration on the surface of the CD4 T lymphocytes compared to that on cell lines used for the neutralization assays could be a contributing factor to the observed differences in neutralizing activity.
The calculation of the Auger line shape is based on a model similar to the one used for Auger neutralization of ions on solid surfaces14.
Laboratory-grade basic materials, namely calcium carbonate (CaCO3), calcium hydroxide (Ca(OH 2), calcium oxide (CaO), sodium carbonate (Na2CO3), sodium hydroxide (NaOH), and ammonia (NH3) were used for acid neutralization.
Two-hundred TCID50 of pseudoviruses were used for the neutralization assay.
Na and Cl ions in the concentration of 0.145 M NaCl model are used for system neutralization.
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