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Anti human fcr blocking reagent

Manufactured by Miltenyi Biotec
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The Anti-human FcR Blocking Reagent is a laboratory product designed to block Fc receptors on human cells. It helps to prevent non-specific binding of antibodies, thus improving the accuracy and reliability of immunoassays and flow cytometry experiments.

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Lab products found in correlation

Anti cd3 apc cy7 clone hit3a, by BioLegend (2 mentions) Anti cd4 apc clone rpa ta, by BioLegend (2 mentions) Anti cd8 percp cy5.5 clone sk1, by BioLegend (2 mentions) Anti ccr4 pe cy7 clone l291h4, by BioLegend (2 mentions) Anti ccr7 fitc clone g043h7, by BioLegend (2 mentions) Anti ccr10 pe clone 314305, by R&D Systems (2 mentions) Anti cd45ro buv395 clone uchl1, by BD (2 mentions) Anti cd3 apc cy7, by BioLegend (2 mentions) Anti cd4 pe cy7 rpa ta, by BioLegend (2 mentions) Anti cd8 bv421 clone rpa t8, by BioLegend (2 mentions) Lsr ll, by BD (2 mentions) Zombie aqua fixable viability kit, by BioLegend (2 mentions) Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (1 mentions) Facsdiva version 6, by BD (1 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions) Lsrii flow cytometer, by BD (1 mentions) Fixable viability dye, by Thermo Fisher Scientific (1 mentions) Tryple, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Human FcR blocking reagent Anti-FcR blocking reagent FcR blocking reagent Human FcR blocking FcR blocking Human blocking reagent Blocking reagent

4 protocols using anti human fcr blocking reagent

1

Multiparameter Phenotyping of Human PBMC

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Freshly isolated or cultured PBMCs were stained with Zombie Aqua™ Fixable Viability Kit (BioLegend) for 20 min at room temperature and were incubated with anti-human FcR Blocking Reagent (Miltenyi Biotec, #130–059-901) and were then incubated with cell surface antibodies for 30 min on ice. The following monoclonal antibodies were used for phenotyping of freshly isolated PBMC: anti-CD3-APC-Cy7 (clone HIT3a, Biolegend, #300317), anti-CD4-APC (clone RPA-TA, Biolegend, #300514), anti-CD8- PerCP-Cy5.5 (clone SK1, Biolegend, #344709), anti-CCR4-PE-Cy7 (clone L291H4, Biolegend, #359409), anti-CCR7-FITC (clone G043H7, Biolegend, #353215), anti-CCR10-PE (clone 314305, R&D #FAB3478P-025), anti-CD45RO-BUV395 (clone UCHL1, BD Biosciences, #564292). And the following monoclonal antibodies were used for analyzing of proliferating lymphocytes within cultured PBMC: anti-CD3-APC-Cy7, anti-CD4-PE-Cy7 (RPA-TA, Biolegend, #300512), anti-CD8-BV421 (clone RPA-T8, Biolegend, #301036). Cells were acquired with an LSR ll (BD Biosciences) and data were analyzed by using FlowJo software (FlowJo, LLC, Ashland, OR, USA).
Kim D., Kobayashi T., Voisin B., Jo J.H., Sakamoto K., Jin S.P., Kelly M., Pasieka H.B., Naff J.L., Meyerle J.H., Ikpeama I.D., Fahle G.A., Davis F.P., Rosenzweig S.D., Alejo J.C., Pittaluga S., Kong H.H., Freeman A.F, & Nagao K. (2020). Targeted therapy guided by single-cell transcriptomic analysis in drug-induced hypersensitivity syndrome/drug reaction with eosinophilia and systemic symptoms: A case report. Nature medicine, 26(2), 236-243.
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2

Multiparameter Phenotyping of Human PBMC

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Freshly isolated or cultured PBMCs were stained with Zombie Aqua™ Fixable Viability Kit (BioLegend) for 20 min at room temperature and were incubated with anti-human FcR Blocking Reagent (Miltenyi Biotec, #130–059-901) and were then incubated with cell surface antibodies for 30 min on ice. The following monoclonal antibodies were used for phenotyping of freshly isolated PBMC: anti-CD3-APC-Cy7 (clone HIT3a, Biolegend, #300317), anti-CD4-APC (clone RPA-TA, Biolegend, #300514), anti-CD8- PerCP-Cy5.5 (clone SK1, Biolegend, #344709), anti-CCR4-PE-Cy7 (clone L291H4, Biolegend, #359409), anti-CCR7-FITC (clone G043H7, Biolegend, #353215), anti-CCR10-PE (clone 314305, R&D #FAB3478P-025), anti-CD45RO-BUV395 (clone UCHL1, BD Biosciences, #564292). And the following monoclonal antibodies were used for analyzing of proliferating lymphocytes within cultured PBMC: anti-CD3-APC-Cy7, anti-CD4-PE-Cy7 (RPA-TA, Biolegend, #300512), anti-CD8-BV421 (clone RPA-T8, Biolegend, #301036). Cells were acquired with an LSR ll (BD Biosciences) and data were analyzed by using FlowJo software (FlowJo, LLC, Ashland, OR, USA).
Kim D., Kobayashi T., Voisin B., Jo J.H., Sakamoto K., Jin S.P., Kelly M., Pasieka H.B., Naff J.L., Meyerle J.H., Ikpeama I.D., Fahle G.A., Davis F.P., Rosenzweig S.D., Alejo J.C., Pittaluga S., Kong H.H., Freeman A.F, & Nagao K. (2020). Targeted therapy guided by single-cell transcriptomic analysis in drug-induced hypersensitivity syndrome/drug reaction with eosinophilia and systemic symptoms: A case report. Nature medicine, 26(2), 236-243.
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3

Flow Cytometry Analysis of Cell Populations

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Cells were examined using flow cytometry (LSRII flow cytometer, BD Biosciences) and data were analyzed with FACSDiva Version 6.1.2 (BD Biosciences) and/or FlowJo_V10 (Ashland, OR, U.S.A) software. Utilized antibodies are listed in Table 3. Before staining, cells were blocked with anti-mouse FcγRII/III (unconjugated, clone 2.4G2, kindly provided by Dr J. Unkeless, Mount Sinai School of Medicine, New York, NY) and anti-human FcR blocking reagent (Miltenyi Biotec). Propidium iodide or the LIVE/DEAD Fixable Aqua Dead Cell Stain Kit (Life Technologies) was used to discriminate live and dead cells. FoxP3/Transcription Factor Staining buffer set (eBioscience, Thermo Fisher Scientific, Waltham, MA) was used to perform transcription factor staining.
To determine absolute cell numbers of HPC or NK cell subpopulations, cultured cells were harvested and counted with trypan blue in a Bürker counting chamber to determine the total number of viable cells. These counted viable cell numbers were normalized to a starting cell number of 1000 cells on day 0 for each condition. FACS analysis was used to determine the percentages of the different subpopulations and the normalized viable cell numbers were then multiplied by the corresponding percentage of each subpopulation.
Kiekens L., Van Loocke W., Taveirne S., Wahlen S., Persyn E., Van Ammel E., De Vos Z., Matthys P., Van Nieuwerburgh F., Taghon T., Van Vlierberghe P., Vandekerckhove B, & Leclercq G. (2021). T-BET and EOMES Accelerate and Enhance Functional Differentiation of Human Natural Killer Cells. Frontiers in Immunology, 12, 732511.
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4

Endothelial Activation Cytokine Profiling

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Confluent endothelial cells were incubated with TDB‐conditioned PBMC supernatant for 6 h at 37°C, 5% CO2. Supernatants were harvested and cytokine analysis was performed. Cells were harvested using TrypLE™ (ThermoFisher Scientific, ref: 12604013), and single‐cell suspensions were incubated 15 min at room temperature in the dark with the fixable viability dye (ref: 65‐0865‐14, ThermoFisher Scientific). After washing with FACS buffer, cells were blocked with anti‐human FcR blocking reagent (Miltenyi Biotec, ref: 130‐059‐901). After blocking, cells were incubated with either mouse anti‐human CD54‐BV421 (BD Biosciences, ref: 566262, clone HA58) or mouse anti‐human CD106‐FITC (BD Biosciences, ref: 551146, clone 51‐10C9). After wash, samples were fixed and run on analyzers.
Himmels P., Nguyen T.T., Mitzner M.C., Arrazate A., Yeung S., Burton J., Clark R., Totpal K., Jesudason R., Yang A., Solon M., Eastham J., Modrusan Z., Webster J.D., Lo A.A., Piskol R, & Ye W. (2023). T cell‐dependent bispecific antibodies alter organ‐specific endothelial cell–T cell interaction. EMBO Reports, 24(3), e55532.
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