Artifacts Arising from Using Leukocytic Fc Receptor Blocking Buffer

• Published in: Bulletin of experimental biology and medicine Vol. 162; no. 2; pp. 244 - 247
• Main Authors: Zubova, S. V., Kabanov, D. S., Serov, D. A., Grachev, S. V., Prokhorenko, I. R.
• Format: Journal Article
• Language: English
• Published: New York Springer US 01-12-2016; Springer; Springer Nature B.V
• Subjects: Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - immunology; B cells; Biomedical and Life Sciences; Biomedicine; Blood cells; Blood Cells - immunology; Blood Cells - metabolism; Buffers; Cell Biology; Dendritic cells; Enzyme-Linked Immunosorbent Assay - standards; Fc receptors; Flow Cytometry - standards; Gene Expression; Humans; Immunoglobulin G; Immunoglobulin G - chemistry; Immunoglobulin G - immunology; Indicators and Reagents - chemistry; Internal Medicine; Laboratory Medicine; Monoclonal antibodies; Monocytes; Monocytes - immunology; Monocytes - metabolism; NF-kappa B p50 Subunit - analysis; NF-kappa B p50 Subunit - genetics; NF-kappa B p50 Subunit - immunology; NF-κB protein; Pathology; Phosphoproteins; Primary Cell Culture; Receptors, Fc - antagonists & inhibitors; Receptors, Fc - genetics; Receptors, Fc - immunology; TLR4 protein; Toll-Like Receptor 4 - analysis; Toll-Like Receptor 4 - genetics; Toll-Like Receptor 4 - immunology; Toll-like receptors; Tumor necrosis factor; Tumor Necrosis Factor-alpha - analysis; Tumor Necrosis Factor-alpha - genetics; Tumor Necrosis Factor-alpha - immunology; Human TruStain FcX; Fcγ receptors; TNF-α; HTA125; NF-κB p50
• ISSN: 0007-4888; 1573-8221
• DOI: 10.1007/s10517-016-3586-9

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.