Fc receptor blocking antibody

Manufactured by R&D Systems

Sourced in United States

The Fc receptor blocking antibody is a laboratory reagent used in immunological research. It functions to block the binding of the Fc region of antibodies to Fc receptors, which can interfere with certain experimental protocols. This product is designed for use in in vitro applications.

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Lab products found in correlation

Lsrii flow cytometer, by BD (6 mentions) Ionomycin, by Merck Group (4 mentions) Golgistop, by BD (4 mentions) Phorbol 12 myristate 13 acetate pma, by Merck Group (3 mentions) Appropriate buffers, by Thermo Fisher Scientific (2 mentions) Anti mouse cd45 pe cy7, by Thermo Fisher Scientific (1 mentions) Flow cytometry antibodies, by BioLegend (1 mentions) Anti ifn γ xmg1, by BioLegend (1 mentions) M5 114, by BioLegend (1 mentions) Anti cd4 rm4 5, by BioLegend (1 mentions) Ctla 4 uc10 4b9, by Thermo Fisher Scientific (1 mentions) Cd206, by Thermo Fisher Scientific (1 mentions) Foxp3 transcription factor staining buffer set, by Thermo Fisher Scientific (1 mentions)

6 protocols using fc receptor blocking antibody

Corneal Cell Phenotyping by Flow Cytometry

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Transplanted corneas were collected and digested with collagenase and Dnase, and single-cell suspensions were prepared and filtered. Cells were incubated with an Fc receptor blocking antibody (R&D Systems, Minneapolis, MN, USA). After they were washed, the cells were incubated with flow cytometry antibodies from BioLegend (San Diego, CA, USA) or Thermo Fisher Scientific for 1 hour: PE-Cy7-anti-mouse CD45; Pacific Blue-anti-mouse CD11b Ab; FITC-anti-mouse CD4 Ab; APC-anti-mouse Ly-6G Ab; and PE-anti-mouse F4/80 Ab. Stained cells were rinsed and analyzed using an LSR II flow cytometer (BD Biosciences, Franklin Lakes, NJ, USA), and the results were analyzed using Summit 4.3 (Agilent Dako, Santa Clara, CA, USA). Dead cells were excluded by performing forward-scatter versus side-scatter analysis, and samples were subsequently analyzed by gating on CD45⁺ cells (Supplementary Fig. S1).

Wang S., Mittal S.K., Lee S., Herrera A.E., Krauthammer M., Elbasiony E., Blanco T., Alemi H., Nakagawa H., Chauhan S.K., Dana R, & Dohlman T.H. (2024). Effector T Cells Promote Fibrosis in Corneal Transplantation Failure. Investigative Ophthalmology & Visual Science, 65(1), 40.

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Immune Cell Profiling by Flow Cytometry

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All cell suspensions were incubated with an Fc receptor blocking antibody (R&D Systems, Minneapolis, MN) for 10 minutes to avoid nonspecific staining, and were subsequently stained with the following antibodies: anti-CD11b (M1/70), antimouse I-A/I-E (major histocompatibility complex (MHC) II, M5/114.15.2), anti-CD4 (RM4–5) and anti-IFNγ (XMG1.2) (Biolegend, San Diego, CA). For intracellular IFNγ staining, cells were stimulated with phorbol 12-myristate 13-acetate (PMA; 50 ng/mL; Sigma-Aldrich) and Ionomycin (500 ng/mL; Sigma-Aldrich) overnight in the presence of Golgistop (0.7 µL/100 µL media; BD Biosciences, San Jose, CA), and were fixed and permeabilized with appropriate buffers (eBioscience). Proper isotype controls were used for all antibodies. Stained cells were analyzed using the LSRII flow cytometer (BD Biosciences, Franklin Lakes, NJ) and the results were analyzed using FlowJo software (FlowJo LLC, Ashland, OR).

Shao C., Chen Y., Nakao T., Amouzegar A., Yin J., Tahvildari M., Lužnik Z., Chauhan S.K, & Dana R. (2019). Local delivery of regulatory T cells promotes corneal allograft survival. Transplantation, 103(1), 182-190.

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Flow Cytometric Analysis of T Cell Subsets

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All cell suspensions were incubated with an Fc receptor blocking antibody (R&D Systems, Minneapolis, MN) before they were stained with the following antibodies: anti-CD45 (30-F11), anti-CD25 (PC61.5), anti-Foxp3 (FJK-16s), and anti-cytotoxic T lymphocyte antigen-4 (CTLA-4) (UC10-4B9) from eBioscience (San Diego, CA); and anti-CD4 (RM4-5) and anti-IFN-γ (XMG1.2) from Biolegend (San Diego, CA). In order to stain intracellular IFN-γ expression, cells were stimulated with phorbol 12-myristate 13-acetate (PMA; 50 ng/ml; Sigma-Aldrich) and Ionomycin (500 ng/ml; Sigma-Aldrich) for 6 hours in the presence of Golgistop (0.7 μl/100 μl media; BD Biosciences, San Jose, CA). For intracellular staining of IFN-γ and intranuclear staining of Foxp3, cells were fixed and permeabilized with appropriate buffers (eBioscience). Isotype controls were used for all antibodies. Cells were analyzed using the LSRII flow cytometer (BD Biosciences, Franklin Lakes, NJ) and Summit v4.3 Software (DAKO Colorado Inc., Fort Collins, CO).

Tahvildari M., Omoto M., Chen Y., Emami-Naeini P., Inomata T., Dohlman T.H., Kaye A.E., Chauhan S.K, & Dana R. (2016). In vivo expansion of regulatory T cells by low-dose interleukin-2 treatment increases allograft survival in corneal transplantation. Transplantation, 100(3), 525-532.

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Flow Cytometric Analysis of Corneal and Conjunctival Immune Cells

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Each group consisted of 3 samples, and every sample contained 4 ipsilateral corneas. Pooled corneas and conjunctiva were used for flow cytometry. All experiments were performed with three repeats. Fc receptor–blocking antibody (R&D Systems, Minneapolis, MN, USA) was used to incubate cell suspension for 10 minutes to avoid nonspecific staining. The cells were subsequently stained with the fluorescence-conjugated antibodies against CD4 (Peprotech), CD25 (Peprotech), Foxp3 (Peprotech), CD45 (Peprotech), F4/80 (Peprotech), CD86 (Peprotech), CD206 (Peprotech), and Areg (Biorbyt). Proper isotype controls were applied for every antibody. Foxp3/transcription factor staining buffer set (Invitrogen) was used for Foxp3 staining. The cells were stained according to the protocol of the set. An LSRII flow cytometer (BD Biosciences) was used to analyze stained cells. The results were analyzed using FlowJo software (FlowJo LLC, Ashland, OR, USA).

Yan D., Yu F., Chen L., Yao Q., Yan C., Zhang S., Wu N., Gong D., Sun H., Fu Y, & Shao C. (2020). Subconjunctival Injection of Regulatory T Cells Potentiates Corneal Healing Via Orchestrating Inflammation and Tissue Repair After Acute Alkali Burn. Investigative Ophthalmology & Visual Science, 61(14), 22.

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Flow Cytometry-Based Immune Cell Profiling

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Cell suspensions were incubated in PBS with zombie death/live (Table 1) for 20 minutes, washed with 10% fetal bovine serum, and incubated with an Fc receptor-blocking antibody (R&D Systems, Minneapolis, MN) 20 minutes. The cells were stained with the fluorescent conjugated antibodies or isotype controls (Table 1). The stained cells were analyzed using an LSRII flow cytometer (BD Biosciences Inc., San Jose, CA), and the acquired data were analyzed using FlowJo software (FlowJo LLC, Ashland, OR). For staining of IFN-γ, cells were stimulated with phorbol 12-myristate 13-acetate (PMA; 50 ng/mL; Sigma Aldrich Corp., St. Louis, MO) and Ionomycin (500 ng/mL; Sigma Aldrich Corp., St. Louis, MO) in the presence of Golgistop (0.7 µL/100 µL media; BD Biosciences Inc., San Jose, CA). Cells were fixed and permeabilized for intracellular staining with appropriate buffers (eBioscience/Thermofisher Inc., San Diego, CA).

van Haastregt J.C., Diederiks J.P., van Rossum E., de Witte L.P., Voorhoeve P.M, & Crebolder H.F. (2000). Effects of a programme of multifactorial home visits on falls and mobility impairments in elderly people at risk: randomised controlled trial. BMJ : British Medical Journal, 321(7267), 994-998.

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Flow Cytometry-Based Cell Phenotyping

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Cell suspensions were incubated in PBS with zombie death/live (Table 1) for 20 minutes, then washed with 10% FBS, incubated with an Fc receptor blocking antibody (R&D Systems, Minneapolis, MN) for 20 minutes, and subsequently stained with the requisite antibodies or isotype controls (Table 1). Stained cells were analyzed using an LSRII flow cytometer (BD Biosciences Inc., San Jose, CA), and the acquired data were analyzed using FlowJo software (FlowJo LLC, Ashland, OR). For staining of IFN-γ, cells were stimulated with phorbol 12-myristate 13-acetate (PMA; 50 ng/mL; Sigma Aldrich Corp., St. Louis, MO) and Ionomycin (500 ng/mL; Sigma Aldrich Corp., St. Louis, MO) in the presence of Golgistop (0.7 μL/100 μL media; BD Biosciences Inc., San Jose, CA). For intracellular staining, cells were fixed and permeabilized with appropriate buffers (eBioscience/Thermofisher Inc., San Diego, CA).

Blanco T., Singh R.B., Nakagawa H., Taketani Y., Dohlman T.H., Chen Y., Chauhan S.K., Yin J, & Dana R. (2023). Conventional Type I Migratory CD103+ Dendritic Cells are Required for Corneal Allograft Survival. Mucosal immunology, 16(5), 711-726.

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