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.
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References
Daëron M. Fc receptor biology. Annu. Rev. Immunol. 1997;15: 203-234.
Dunn-Siegrist I, Leger O, Daubeuf B, Poitevin Y, Dépis F, Herren S, Kosco-Vilbois M, Dean Y, Pugin J, Elson G. Pivotal involvement of Fcgamma receptor IIA in the neutralization of lipopolysaccharide signaling via a potent novel anti-TLR4 monoclonal antibody 15C1. J. Biol. Chem. 2007; 282(48):34,817-34,827.
Flo TH, Ryan L, Latz E, Takeuchi O, Monks BG, Lien E, Halaas O, Akira S, Skjåk-Braek G, Golenbock DT, Espevik T. Involvement of toll-like receptor (TLR) 2 and TLR4 in cell activation by mannuronic acid polymers. J. Biol. Chem. 2002;277(38):35 489-35 495.
Jitschin R, Hofmann AD, Bruns H, Giessl A, Bricks J, Berger J, Saul D, Eckart MJ, Mackensen A, Mougiakakos D. Mitochondrial metabolism contributes to oxidative stress and reveals therapeutic targets in chronic lymphocytic leukemia. Blood. 2014;123(17):2663-2672.
Klingenberg R, Brokopp CE, Grivès A, Courtier A, Jaguszewski M, Pasqual N, Vlaskou Badra E, Lewandowski A, Gaemperli O, Hoerstrup SP, Maier W, Landmesser U, Lüscher TF, Matter CM. Clonal restriction and predominance of regulatory T cells in coronary thrombi of patients with acute coronary syndromes. Eur. Heart J. 2015;36(17):1041-1048.
Lien E, Means TK, Heine H, Yoshimura A, Kusumoto S, Fukase K, Fenton MJ, Oikawa M, Qureshi N, Monks B, Finberg RW, Ingalls RR, Golenbock DT. Toll-like receptor 4 imparts ligand-specific recognition of bacterial lipopolysaccharide. J. Clin. Invest. 2000;105(4):497-504.
Muroi M, Tanamoto K. The polysaccharide portion plays an indispensable role in Salmonella lipopolysaccharide-induced activation of NF-kappaB through human toll-like receptor 4. Infect. Immun. 2002;70(11):6043-6047.
Nimmerjahn F, Ravetch JV. Fcgamma receptors as regulators of immune responses. Nat. Rev. Immunol. 2008;8(1):34-47.
van de Winkel JG, Anderson CL. Biology of human immunoglobulin G Fc receptors. J. Leukoc. Biol. 1991;49(5):511-524.
Vogelpoel LT, Baeten DL, de Jong EC, den Dunnen J. Control of cytokine production by human fc gamma receptors: implications for pathogen defense and autoimmunity. Front. Immunol. 2015;6:79. doi: 10.3389/fimmu.2015.00079.
Wang R, Stephens J, Lacy MJ. Characterization of monoclonal antibody HTA125 with specificity for human TLR4. Hybrid. Hybridomics. 2003;22(6):357-365.
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Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 162, No. 8, pp. 210-213, August, 2016
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Zubova, S.V., Kabanov, D.S., Serov, D.A. et al. Artifacts Arising from Using Leukocytic Fc Receptor Blocking Buffer. Bull Exp Biol Med 162, 244–247 (2016). https://doi.org/10.1007/s10517-016-3586-9
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DOI: https://doi.org/10.1007/s10517-016-3586-9