Abstract
We studied the effects of Human TruStain FcX buffer (Fcγ receptor blocking solution) in experiments on evaluation of TLR4 level with labeled monoclonal antibodies, intracellular immunofluorescent staining of NF-κB p50, and TNF-α synthesis on human isolated monocytes and whole blood cells. The influence of the blocking buffer on the measured parameters should be taken into account and appropriateness of its use in experiments on isolated cells and whole blood should be considered.
Keywords:
Fcγ receptors; HTA125; Human TruStain FcX; NF-κB p50; TNF-α.
MeSH terms
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Antibodies, Monoclonal / chemistry
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Antibodies, Monoclonal / immunology
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Artifacts*
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Blood Cells / immunology
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Blood Cells / metabolism
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Buffers
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Enzyme-Linked Immunosorbent Assay / standards
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Flow Cytometry / standards
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Gene Expression
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Humans
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Immunoglobulin G / chemistry
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Immunoglobulin G / immunology
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Indicators and Reagents / chemistry
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Monocytes / immunology
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Monocytes / metabolism*
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NF-kappa B p50 Subunit / analysis*
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NF-kappa B p50 Subunit / genetics
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NF-kappa B p50 Subunit / immunology
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Primary Cell Culture
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Receptors, Fc / antagonists & inhibitors*
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Receptors, Fc / genetics
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Receptors, Fc / immunology
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Toll-Like Receptor 4 / analysis*
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Toll-Like Receptor 4 / genetics
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Toll-Like Receptor 4 / immunology
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Tumor Necrosis Factor-alpha / analysis*
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Tumor Necrosis Factor-alpha / genetics
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Tumor Necrosis Factor-alpha / immunology
Substances
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Antibodies, Monoclonal
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Buffers
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Immunoglobulin G
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Indicators and Reagents
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NF-kappa B p50 Subunit
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Receptors, Fc
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TLR4 protein, human
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Toll-Like Receptor 4
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Tumor Necrosis Factor-alpha