Fc receptor block

Manufactured by BioLegend

Sourced in United States

The Fc receptor block is a reagent designed to prevent non-specific binding of antibodies to Fc receptors. It helps reduce background and increase the specificity of immunoassays and flow cytometry experiments.

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Lab products found in correlation

Aurora flow cytometer, by Cytek (2 mentions) Zombie nir fixable viability dye, by BioLegend (2 mentions) Facscanto 2, by BD (1 mentions) Mitotracker green, by Thermo Fisher Scientific (1 mentions) 40 m cell strainer, by BD (1 mentions) Flow cytometry staining buffer, by Thermo Fisher Scientific (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) Apc anti human cd11b, by BioLegend (1 mentions) Fitc anti human cd206, by BioLegend (1 mentions) Pe anti human cd80, by BioLegend (1 mentions) Cd11b fitc, by BioLegend (1 mentions) Ly6g apc, by BioLegend (1 mentions) Gentlemacs dissociator, by Miltenyi Biotec (1 mentions) Cd45 bv711, by BioLegend (1 mentions) Intercalator ir, by Standard BioTools (1 mentions) Maxpar pbs, by Standard BioTools (1 mentions) Facscanto, by BD (1 mentions)

Close spelling variants of this product

Fc block (182 citations) TruStain FcX Fc receptor block (5 citations) Human Fc receptor block (2 citations) TruStain Fc receptor block (2 citations)

8 protocols using fc receptor block

Multiparameter Flow Cytometry Profiling

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Isolated PBMCs (2 × 10⁶) were centrifuged and resuspended in 75 μl FACS buffer (phosphate-buffered saline (PBS) and 2% fetal bovine serum (FBS)) and 5 μl Fc receptor block (BioLegend, no. 422302) for 5 min at room temperature. For samples stained with anti-IgG, it was observed that Fc block inappropriately interfered with staining, so a preincubation step of the anti-IgG alone for 5 min at 22 °C was added before the addition of the block. Next, 25 μl of antibody cocktail (Supplementary Table 3) was added (100 μl staining reaction), and samples were incubated for 20 min at 4 °C. Cells were washed in PBS and resuspended in a PBS dilution of Zombie NIR fixable viability dye (BioLegend, no. 423106). Cells were washed and fixed in 0.8% paraformaldehyde for 10 min at 22 °C in the dark before a final wash and resuspension for analysis.
Cells were analysed on a Cytek Aurora flow cytometer using Cytek SpectroFlo software. Up to 3 × 10⁶ cells were analysed using FlowJo v.10 (Treestar) software.

Woodruff M.C., Ramonell R.P., Haddad N.S., Anam F.A., Rudolph M.E., Walker T.A., Truong A.D., Dixit A.N., Han J.E., Cabrera-Mora M., Runnstrom M.C., Bugrovsky R., Hom J., Connolly E.C., Albizua I., Javia V., Cashman K.S., Nguyen D.C., Kyu S., Singh Saini A., Piazza M., Tipton C.M., Khosroshahi A., Gibson G., Martin G.S., Maier C.L., Esper A., Jenks S.A., Lee F.E, & Sanz I. (2022). Dysregulated naive B cells and de novo autoreactivity in severe COVID-19. Nature, 611(7934), 139-147.

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Osteoclast Galectin Expression Profiling

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Single-cell suspensions of osteoclasts were collected after passing through a 40 µm cell strainer (BD Bioscience), incubated with Fc receptor block (#101319; Biolegend) for 10 min, and then incubated with CoraLite 488–conjugated galectin-3 polyclonal antibody (rabbit anti-mouse antibody; CL488-14979; Proteintech; epitopes mapped throughout the full-length protein), eFluor 660–conjugated anti–galectin-3 monoclonal antibody (rabbit anti-mouse antibody; #50-5301-82; Thermo Fisher Scientific; clone M3/38, epitopes mapped within the N-terminal region), PE-conjugated anti–galectin-1 antibody (goat anti-mouse antibody; IC1245P; R&D), or the corresponding rabbit and goat isotype control (#31235; #31245; Invitrogen) in flow cytometry staining buffer (eBioscience) for 30 min at 4°C. For Mitotracker Green staining, cells were incubated with 100 nM Mitotracker Green (M7514; Invitrogen) in HBSS for 45 min. Then, cells were subjected to flow cytometry analysis on a FACS Canto II (BD Bioscience). Data analysis was carried out using FlowJo software.

Zhu L., Tang Y., Li X.Y., Kerk S.A., Lyssiotis C.A., Sun X., Wang Z., Cho J.S., Ma J, & Weiss S.J. (2023). Proteolytic regulation of a galectin-3/Lrp1 axis controls osteoclast-mediated bone resorption. The Journal of Cell Biology, 222(4), e202206121.

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Phenotyping M1 and M2 Macrophages

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Flow cytometry was performed using standard protocols. An Fc receptor block (BioLegend) was incubated with samples to reduce nonspecific antibody binding for half an hour at 4 °C. Afterward, the cells were incubated for half an hour at 4 °C with fluorochrome-tagged monoclonal antibodies from BioLegend, such as anti-human CD11b APC (cat# 301330), anti-human CD80 PE (cat# 305220), anti-human CD11c APC (cat# 301614), and anti-human CD206 FITC (cat#321110). Cell populations were gated as follows: M1 macrophages (CD11b+CD 80+) and M2 macrophages (CD11b+ CD206+). We performed flow cytometry using a FACSCalibur flow cytometer (BD Bioscience), and FlowJo software (FlowJo, LCC) was used for data analysis.

Xue R., Xie M., Wu Z., Wang S., Zhang Y., Han Z., Li C., Tang Q., Wang L., Li D., Wang S., Yang H, & Zhao R.C. (2024). Mesenchymal Stem Cell-Derived Exosomes Promote Recovery of The Facial Nerve Injury through Regulating Macrophage M1 and M2 Polarization by Targeting the P38 MAPK/NF-Κb Pathway. Aging and Disease, 15(2), 851-868.

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Poly-A Particle Toxicity Evaluation

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Select groups of mice received a ‘long-term’ dosage of Poly-A. Mice received a daily tail vein injection of 2 × 10⁸ (~1 µm) Poly-A particles in 100 µL sterile phosphate-buffered saline (PBS). Control mice received either an equivalent amount of PBS (‘saline’) alone or ‘free aspirin’—salicylic acid. Mice were weighed and underwent health scoring daily before euthanasia on the fifth day. At the time of euthanasia, a cardiac puncture blood draw was administered, and the liver was harvested. Blood was immediately placed on ice, blocked with 1 µL FC-receptor block (BioLegend #101320) for 10 min. Samples were then stained with 0.1 µg/mL CD45-BV711 (Biolegend #103147), 0.25 µg/mL CD11b-FITC (Biolegend #101206), and 0.1 µg/mL Ly6G-APC (Biolegend #127614) and lyse/fixed (eBioscience) before using an Attune flow cytometer to analyze circulating leukocyte populations. The liver was added to RPMI media with collagenase and DNAse and dissociated using a gentleMACS Dissociator (Miltenyi Biotec). The dissociated liver was strained, centrifuged, and the supernatant was utilized in an aspartate aminotransferase (AST) activity assay (Sigma Aldrich).

Banka A.L., Guevara M.V., Brannon E.R., Nguyen N.Q., Song S., Cady G., Pinsky D.J., Uhrich K.E., Adili R., Holinstat M, & Eniola-Adefeso O. (2023). Cargo-free particles divert neutrophil-platelet aggregates to reduce thromboinflammation. Nature Communications, 14, 2462.

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Comprehensive Immune Profiling of OOC Cells

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After 24 hours of control or LPS treatment, the media from the devices was removed. All staining procedures were conducted inside the OOC devices. Then cells were treated with Fc-receptor block (Biolegend, San Diego, CA, USA, Cat. No: 101302) for 10 min at room temperature. Then a surface antibody cocktail (CD3, CD4, CD25, CD45, CD8a, HLA-DR, CD11c, CD68, CD80, CD19, CD66b, CD206/MMR, CD56, CD86, CD16) was added and incubated for 30 min at room temperature. Cells were then washed with Maxpar PBS (Fluidigm, San Francisco, CA, USA, Cat. No: 201058) for 5 min at room temperature. Cells were fixed with 4% paraformaldehyde in PBS for 20 min at room temperature. After washing with Maxpar PBS (Fluidigm) for 5 min, an intracellular antibody cocktail (Cytokeratin, Vimentin, IFNg, IL-17A, IL-4) was added to the cells for 1 h at room temperature. Cells were then washed 3 times with Maxpar PBS (Fluidigm) for 5 min and incubated with 125 nM of intercalator-Ir (Fluidigm, Cat. No: 201192A) for 1h at room temperature. Lastly, cells were washed 3 times with Maxpar PBS (Fluidigm) for 5 min and once with Maxpar H₂O (Fluidigm, Cat. No: 201069). After staining was complete, the PDMS chamber was removed, and the adherent stained cells that remained on the glass side were air dried for Hyperion ablation.

Reeder J.C., Cowman A.F., Davern K.M., Beeson J.G., Thompson J.K., Rogerson S.J, & Brown G.V. (1999). The adhesion of Plasmodium falciparum-infected erythrocytes to chondroitin sulfate A is mediated by P. falciparum erythrocyte membrane protein 1. Proceedings of the National Academy of Sciences of the United States of America, 96(9), 5198-5202.

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Neutrophil Activation by Chitobiose and β-Glucan

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Neutrophils were blocked in ice cold DPBS containing Fc receptor block (BioLegend, San Diego, CA) for 15 min on ice. Cells were treated with chitobiose (5 mM) or β-glucan (3 mg/mL) for 5 min then stained with 5 μg/mL FITC-labeled mAb for 10 min on ice. Cells were then washed twice resuspended in ice cold 10% buffered formaldehyde. Analysis was performed on a MACSQuant flow cytometer (Miltenyi Biotec, Bergisch Gladbach, Germany) and analyzed using FlowJo software (Tree Star, Ashland, OR) and gated on neutrophils.

Schmitz N.A., Thakare R.P., Chung C.S., Lee C.M., Elias J.A., Lee C.G, & LeBlanc B.W. (2021). Chitotriosidase Activity Is Counterproductive in a Mouse Model of Systemic Candidiasis. Frontiers in Immunology, 12, 626798.

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Multiparametric Flow Cytometry of PBMCs

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Isolated PBMCs (10 × 10⁶) were centrifuged and resuspended in 75 μl FACS buffer (PBS + 2% FBS) and 5 μl Fc receptor block (BioLegend, no. 422302) for 5 min at room temperature. For samples stained with anti-IgG, it was observed that Fc block inappropriately interfered with staining, so a preincubation step of the anti-IgG alone for 5 min at 22 °C was added before the addition of the block. Next, 25 μl of antibody cocktail (Supplementary Table 1) was added (100 μl staining reaction), and samples were incubated for 20 min at 4 °C. Cells were washed in PBS, and resuspended in a PBS dilution of Zombie NIR fixable viability dye (BioLegend, no. 423106). Cells were washed and fixed at 0.8% paraformaldehyde (PFA) for 10 min at 22 °C in the dark before a final wash and resuspension for analysis.
Cells were analyzed on a Cytek Aurora flow cytometer using Cytek SpectroFlo software. Up to 3 × 10⁶ cells were analyzed using FlowJo v10 (Treestar).

Woodruff M.C., Bonham K.S., Anam F.A., Walker T.A., Faliti C.E., Ishii Y., Kaminski C.Y., Ruunstrom M.C., Cooper K.R., Truong A.D., Dixit A.N., Han J.E., Ramonell R.P., Haddad N.S., Rudolph M.E., Yalavarthi S., Betin V., Natoli T., Navaz S., Jenks S.A., Zuo Y., Knight J.S., Khosroshahi A., Lee F.E, & Sanz I. (2023). Chronic inflammation, neutrophil activity, and autoreactivity splits long COVID. Nature Communications, 14, 4201.

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Immunostaining of non-permeabilized cells

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Non-permeabilized cells were stained with the primary antibody clones in FACS buffer (HBSS without Ca²⁺/Mg2⁺, FBS [3%], NaN₃ [0.02%], and CaCl₂ [2.5 mM]), then washed and stained with fluorescently conjugated secondary antibody (goat anti-mouse; catalog no. 115-006-003; Jackson ImmunoResearch). Stained cells were then fixed in 1% paraformaldehyde. Splenocytes were pre-treated with Fc receptor block (catalog no. 422301; BioLegend). PAG-GFP-expressing cells were fixed in 1% paraformaldehyde. Events were recorded using FACSCanto (BD) and analyzed using FlowJo software (version 10.8.1).

Strazza M., Moore E.K., Adam K., Azoulay-Alfaguter I, & Mor A. (2022). Neutralization of the adaptor protein PAG by monoclonal antibody limits murine tumor growth. Molecular Therapy. Methods & Clinical Development, 27, 380-390.

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